Pyrimidine derivatives used as PI-3-kinase inhibitors

ABSTRACT

Phosphatidylinositol (PI) 3-kinase inhibitor compounds (I), their pharmaceutically acceptable salts, and prodrugs thereof; compositions of the new compounds, either alone or in combination with at least one additional therapeutic agent, with a pharmaceutically acceptable carrier; and uses of the new compounds, either alone or in combination with at least one additional therapeutic agent, in the prophylaxis or treatment of proliferative diseases characterized by the abnormal activity of growth factors, protein serine/threonine kinases, and phospholipid kinases.

FIELD OF THE INVENTION

The present invention relates to new phosphatidylinositol (PI) 3-kinaseinhibitor compounds, their pharmaceutically acceptable salts, andprodrugs thereof; compositions of the new compounds, either alone or incombination with at least one additional therapeutic agent, with apharmaceutically acceptable carrier; and uses of the new compounds,either alone or in combination with at least one additional therapeuticagent, in the prophylaxis or treatment of a number of diseases, inparticular, those characterized by the abnormal activity of growthfactors, receptor tyrosine kinases, protein serine/threonine kinases, Gprotein coupled receptors and phospholipid kinases and phosphatases.

BACKGROUND OF THE INVENTION

Phosphatidylinositol 3-kinases (PI3Ks) comprise a family of lipid andserine/threonine kinases that catalyze the transfer of phosphate to theD-3′ position of inositol lipids to produce phosphoinositol-3-phosphate(PIP), phosphoinositol-3,4-diphosphate (PIP₂) andphosphoinositol-3,4,5-triphosphate (PIP₃) that, in turn, act as secondmessengers in signaling cascades by docking proteins containingpleckstrin-homology, FYVE, Phox and other phospholipid-binding domainsinto a variety of signaling complexes often at the plasma membrane((Vanhaesebroeck et al., Annu. Rev. Biochem 70:535 (2001); Katso et al.,Annu. Rev. Cell Dev. Biol. 17:615 (2001)). Of the two Class 1 PI3Ks,Class 1A PI3Ks are heterodimers composed of a catalytic p110 subunit (α,β, δ isoforms) constitutively associated with a regulatory subunit thatcan be p85α, p55α, p50α, p85β or p55γ. The Class 1B sub-class has onefamily member, a heterodimer composed of a catalytic p110γ subunitassociated with one of two regulatory subunits, p101 or p84 (Fruman etal., Annu Rev. Biochem. 67:481 (1998); Suire et al., Curr. Biol. 15:566(2005)). The modular domains of the p85/55/50 subunits include SrcHomology (SH2) domains that bind phosphotyrosine residues in a specificsequence context on activated receptor and cytoplasmic tyrosine kinases,resulting in activation and localization of Class 1A PI3Ks. Class 1BPI3K is activated directly by G protein-coupled receptors that bind adiverse repertoire of peptide and non-peptide ligands (Stephens et al.,Cell 89:105 (1997)); Katso et al., Annu. Rev. Cell Dev. Biol. 17:615-675(2001)). Consequently, the resultant phospholipid products of class IPI3K link upstream receptors with downstream cellular activitiesincluding proliferation, survival, chemotaxis, cellular trafficking,motility, metabolism, inflammatory and allergic responses, transcriptionand translation (Cantley et al., Cell 64:281 (1991); Escobedo andWilliams, Nature 335:85 (1988); Fantl et al., Cell 69:413 (1992)).

In many cases, PIP2 and PIP3 recruit Akt, the product of the humanhomologue of the viral oncogene v-Akt, to the plasma membrane where itacts as a nodal point for many intracellular signaling pathwaysimportant for growth and survival (Fantl et al., Cell 69:413-423 (1992);Bader et al., Nat Rev. Cancer 5:921 (2005); Vivanco and Sawyer, Nat.Rev. Cancer 2:489 (2002)). Aberrant regulation of PI3K, which oftenincreases survival through Akt activation, is one of the most prevalentevents in human cancer and has been shown to occur at multiple levels.The tumor suppressor gene PTEN, which dephosphorylates phosphoinositidesat the 3′ position of the inositol ring and in so doing antagonizes PI3Kactivity, is functionally deleted in a variety of tumors. In othertumors, the genes for the p110α isoform, PIK3CA, and for Akt areamplified and increased protein expression of their gene products hasbeen demonstrated in several human cancers. Furthermore, mutations andtranslocation of p85α that serve to up-regulate the p85-p110 complexhave been described in a few human cancers. Finally, somatic missensemutations in PIK3CA that activate downstream signaling pathways havebeen described at significant frequencies in a wide diversity of humancancers (Kang et al., Proc. Natl. Acad. Sci. USA 102:802 (2005); Samuelset al., Science 304:554 (2004); Samuels et al., Cancer Cell 7:561-573(2005)). These observations show that deregulation of phosphoinositol-3kinase and the upstream and downstream components of this signalingpathway is one of the most common deregulations associated with humancancers and proliferative diseases (Parsons et al., Nature 436:792(2005); Hennessey et al., Nature Rev. Drug Dis. 4:988-1004 (2005)).

SUMMARY OF THE INVENTION

The present invention provides new phosphatidylinositol 3-kinase (PI3K)inhibitor compounds, pharmaceutical formulations that include thecompounds, methods of inhibiting phosphatidylinositol 3-kinase (PI3K),and methods of treating proliferative diseases.

In one aspect of the present invention, new phosphatidylinositol3-kinase (PI3K) inhibitor compounds that are pyrimidine-based compounds,their pharmaceutically acceptable salts, and prodrugs thereof areprovided. The pyrimidine compounds, pharmaceutically acceptable salts,and prodrugs are PI3K inhibitors and are useful in the treatment ofcellular proliferative diseases.

One embodiment of the invention provides a compound having Formula I:

or a stereoisomer, tautomer, or pharmaceutically acceptable saltthereof, wherein,

W is CR_(w) or N, wherein R_(w) is selected from the group consisting of

(1) hydrogen,

(2) cyano,

(3) halogen,

(4) methyl,

(5) trifluoromethyl,

(6) sulfonamido;

R₁ is selected from the group consisting of

(1) hydrogen,

(2) cyano,

(3) nitro,

(4) halogen,

(5) substituted and unsubstituted alkyl,

(6) substituted and unsubstituted alkenyl,

(7) substituted and unsubstituted alkynyl,

(8) substituted and unsubstituted aryl,

(9) substituted and unsubstituted heteroaryl,

(10) substituted and unsubstituted heterocyclyl,

(11) substituted and unsubstituted cycloalkyl,

(12) —COR_(1a),

(13) —CO₂R_(1a),

(14) —CONR_(1a)R_(1b),

(15) —NR_(1a)R_(1b),

(16) —NR_(1a)COR_(1b),

(17) —NR_(1a)SO₂R_(1b),

(18) —OCOR_(1a),

(19) —OR_(1a),

(20) —SR_(1a),

(21) —SOR_(1a),

(22) —SO₂R_(1a), and

(23) —SO₂NR_(1a)R_(1b),

wherein R_(1a), and R_(1b) are independently selected from the groupconsisting of

(a) hydrogen,

(b) substituted or unsubstituted alkyl,

(c) substituted and unsubstituted aryl,

(d) substituted and unsubstituted heteroaryl,

(e) substituted and unsubstituted heterocyclyl, and

(f) substituted and unsubstituted cycloalkyl;

R₂ is selected from the group consisting

(1) hydrogen,

(2) cyano,

(3) nitro,

(4) halogen,

(5) hydroxy,

(6) amino,

(7) substituted and unsubstituted alkyl,

(8) —COR_(2a), and

(9) —NR_(2a)COR_(2b),

wherein R_(2a), and R_(2b) are independently selected from the groupconsisting of

(a) hydrogen, and

(b) substituted or unsubstituted alkyl;

R₃ is selected from the group consisting of

(1) hydrogen,

(2) cyano,

(3) nitro,

(4) halogen,

(5) substituted and unsubstituted alkyl,

(6) substituted and unsubstituted alkenyl,

(7) substituted and unsubstituted alkynyl,

(8) substituted and unsubstituted aryl,

(9) substituted and unsubstituted heteroaryl,

(10) substituted and unsubstituted heterocyclyl,

(11) substituted and unsubstituted cycloalkyl,

(12) —COR_(3a),

(13) —NR_(3a)R_(3b),

(14) —NR_(3a)COR_(3b),

(15) —NR_(3a)SO₂R_(3b),

(16) —OR_(3a),

(17) —SR_(3a),

(18) —SOR_(3a),

(19) —SO₂R_(3a), and

(20) —SO₂NR_(3a)R_(3b),

wherein R_(3a), and R_(3b) are independently selected from the groupconsisting of

(a) hydrogen,

(b) substituted or unsubstituted alkyl,

(c) substituted and unsubstituted aryl,

(d) substituted and unsubstituted heteroaryl,

(e) substituted and unsubstituted heterocyclyl, and

(f) substituted and unsubstituted cycloalkyl; and

R₄ is selected from the group consisting of

(1) hydrogen, and

(2) halogen.

In another embodiment thereof, R₁ comprises substituted or unsubstitutedarylalkyl, substituted or unsubstituted heteroarylalkyl, substituted orunsubstituted cycloalkylalkyl, or substituted or unsubstitutedheterocyclylalkyl.

In a more particular embodiment, W is CH.

In another embodiment, W is N. In a more particular embodiment thereof,R₃ is ═O.

In another embodiment, R₁ is selected from the group consisting of

(1) substituted and unsubstituted alkyl,

(2) substituted and unsubstituted aryl,

(3) substituted and unsubstituted heteroaryl,

(4) substituted and unsubstituted heterocyclyl,

(5) substituted and unsubstituted cycloalkyl,

(6) —OR_(1a), and

(7) —NR_(1a)R_(1b),

wherein R_(1a) and R_(1b) are independently selected from the groupconsisting of

(a) substituted and unsubstituted heteroaryl, and

(b) substituted and unsubstituted heterocyclyl.

In another embodiment R₁ is substituted or unsubstituted heterocyclyl,or substituted or unsubstituted —O-heterocyclyl. In another embodiment,R₁ is substituted or unsubstituted morpholinyl; more particular still,R₁ is unsubstituted N-linked morpholinyl.

In another embodiment thereof, R₁ comprises substituted or unsubstitutedheterocyclylalkyl, or substituted or unsubstituted heteroarylalkyl. Inanother embodiment, R₁ comprises substituted or unsubstitutedmorpholinyl; more particular still, morpholinyl comprises N-linkedmorpholinyl.

In another embodiment, R₁ is substituted or unsubstitutedtetrahydropyran or substituted or unsubstituted tetrahydropyranyloxy.More particular still, R₁ is unsubstituted 4-tetrahydropyranyloxy.

In another embodiment thereof, R₁ comprises substituted or unsubstitutedtetrahydropyran. In a more particular embodiment, tetrahydropyrancomprises 4-tetrahydropyranyloxy.

In another embodiment, R₁ is substituted or unsubstitutedtetrahydrofuran or substituted or unsubstituted tetrahydrofuranyloxy.More particular still, R₁ is unsubstituted 3-tetrahydrofuranyloxy.

In another embodiment, R₁ comprises substituted or unsubstitutedtetrahydrofuran. In another embodiment thereof, tetrahydrofurancomprises 3-tetrahydrofuranyloxy.

In another embodiment, R₂ is selected from the group consisting

(1) hydrogen,

(2) cyano,

(3) hydroxy,

(4) halogen,

(5) amino,

(6) methyl, and

(7) trifluoromethyl.

In another embodiment, R₂ is hydrogen or halogen. In a more particularembodiment, R₂ is hydrogen.

In another embodiment, R₃ is selected from the group consisting of

(1) cyano,

(2) nitro,

(3) halogen,

(4) hydroxy,

(5) amino, and

(6) trifluoromethyl.

In another embodiment, R₃ is trifluoromethyl. In another embodiment, R₃is cyano.

Another embodiment of the invention provides a compound having FormulaII:

or a stereoisomer, tautomer, or pharmaceutically acceptable saltthereof, wherein,

W is CR_(w) or N, wherein R_(w) is selected from the group consisting of

(1) hydrogen,

(2) cyano,

(3) halogen,

(4) methyl,

(5) trifluoromethyl, and

(6) sulfonamido;

X is O, S, NH, or a direct bond;

R₂ is selected from the group consisting

(1) hydrogen,

(2) cyano,

(3) nitro,

(4) halogen,

(5) hydroxy,

(6) amino,

(7) substituted and unsubstituted alkyl,

(8) —COR_(2a), and

(9) —NR_(2a)COR_(2b),

wherein R_(2a), and R_(2b) are independently selected from the groupconsisting of

(a) hydrogen, and

(b) substituted or unsubstituted alkyl;

R₃ is selected from the group consisting of

(1) hydrogen,

(2) cyano,

(3) nitro,

(4) halogen,

(5) substituted and unsubstituted alkyl,

(6) substituted and unsubstituted alkenyl,

(7) substituted and unsubstituted alkynyl,

(8) substituted and unsubstituted aryl,

(9) substituted and unsubstituted heteroaryl,

(10) substituted and unsubstituted heterocyclyl,

(11) substituted and unsubstituted cycloalkyl,

(12) —COR_(3a),

(13) —NR_(3a)R_(3b),

(14) —NR_(3a)COR_(3b),

(15) —NR_(3a)SO₂R_(3b),

(16) —OR_(3a),

(17) —SR_(3a),

(18) —SOR_(3a),

(19) —SO₂R_(3a), and

(20) —SO₂NR_(3a)R_(3b),

wherein R_(3a), and R_(3b) are independently selected from the groupconsisting of

(a) hydrogen,

(b) substituted or unsubstituted alkyl,

(c) substituted and unsubstituted aryl,

(d) substituted and unsubstituted heteroaryl,

(e) substituted and unsubstituted heterocyclyl, and

(f) substituted and unsubstituted cycloalkyl;

R₄ is selected from the group consisting of

(1) hydrogen, and

(2) halogen; and

R₅ is selected from the group consisting of

(1) substituted and unsubstituted cycloalkyl,

(2) substituted and unsubstituted heterocyclyl,

(3) substituted and unsubstituted aryl, and

(4) substituted and unsubstituted heteroaryl.

In another embodiment of Formula II, R₂ is selected from the groupconsisting of

(1) hydrogen,

(2) cyano,

(3) hydroxy,

(4) amino,

(5) halogen, and

(6) substituted and unsubstituted C₁₋₃ alkyl.

In another embodiment of Formula II, R₃ is selected from the groupconsisting of

(1) hydrogen,

(2) cyano,

(3) thio,

(4) halogen,

(5) nitro,

(6) substituted and unsubstituted alkyl,

(7) substituted and unsubstituted alkenyl,

(8) substituted and unsubstituted alkynyl,

(9) —OR_(3a),

(10) —NR_(3a)R_(3b),

(11) —COR_(3a), and

(12) —NR_(3a)COR_(3b),

wherein R_(3a), and R_(3b) are independently selected from the groupconsisting of

(a) hydrogen, and

(b) substituted or unsubstituted alkyl.

In another embodiment of Formula II, R₃ is trifluoromethyl. In anotherembodiment, W is CH. In another embodiment, R₂ is H.

In another embodiment of Formula II, R₅ is selected from the groupconsisting of

(1) substituted or unsubstituted morpholinyl,

(2) substituted or unsubstituted tetrahydropyranyl, and

(3) substituted or unsubstituted tetrahydropyranyl.

In a more particular embodiment thereof, R₅ is N-linked morpholinyl;more particular still, X is a direct link. In another more particularembodiment, R₅ is 4-tetrahydropyranyl; more particular still, X is O. Inanother embodiment, R₅ is 3-tetrahydrofuranyl; more particular still, Xis O.

In another embodiment, W is N. In a more particular embodiment thereof,R₃ is ═O.

Another embodiment of the invention provides a compound having FormulaIII:

or a stereoisomer, tautomer, or pharmaceutically acceptable saltthereof, wherein,

W is CR_(w) or N, wherein R_(w) is selected from the group consisting of

(1) hydrogen,

(2) cyano,

(3) halogen,

(4) methyl,

(5) trifluoromethyl, and

(6) sulfonamido;

R₂ is selected from the group consisting

(1) hydrogen,

(2) cyano,

(3) nitro,

(4) halogen,

(5) hydroxy,

(6) amino,

(7) substituted and unsubstituted alkyl,

(8) —COR_(2a), and

(9) —NR_(2a)COR_(2b),

wherein R_(2a), and R_(2b) are independently selected from the groupconsisting of

(a) hydrogen, and

(b) substituted or unsubstituted alkyl;

R₃ is selected from the group consisting of

(1) hydrogen,

(2) cyano,

(3) nitro,

(4) halogen,

(5) substituted and unsubstituted alkyl,

(6) substituted and unsubstituted alkenyl,

(7) substituted and unsubstituted alkynyl,

(8) substituted and unsubstituted aryl,

(9) substituted and unsubstituted heteroaryl,

(10) substituted and unsubstituted heterocyclyl,

(11) substituted and unsubstituted cycloalkyl,

(12) —COR_(3a),

(13) —NR_(3a)R_(3b),

(14) —NR_(3a)COR_(3b),

(15) —NR_(3a)SO₂R_(3b),

(16) —OR_(3a),

(17) —SR_(3a),

(18) —SOR_(3a),

(19) —SO₂R_(3a), and

(20) —SO₂NR_(3a)R_(3b),

wherein R_(3a), and R_(3b) are independently selected from the groupconsisting of

(a) hydrogen,

(b) substituted or unsubstituted alkyl,

(c) substituted and unsubstituted aryl,

(d) substituted and unsubstituted heteroaryl,

(e) substituted and unsubstituted heterocyclyl, and

(f) substituted and unsubstituted cycloalkyl;

R₄ is selected from the group consisting of

(1) hydrogen, and

(2) halogen; and

R₆ is selected from the group consisting of

(1) hydrogen,

(2) substituted and unsubstituted alkyl, and

(3) substituted and unsubstituted cycloalkyl.

In another embodiment of Formula III, R₂ is selected from the groupconsisting

(1) hydrogen,

(2) cyano,

(3) hydroxy,

(4) halogen,

(5) amino,

(6) methyl, and

(7) trifluoromethyl.

In another embodiment of Formula III, R₃ is selected from the groupconsisting of

(1) cyano,

(2) nitro,

(3) halogen,

(4) hydroxy,

(5) amino, and

(6) trifluoromethyl.

In another embodiment of Formula III, R₆ is selected from the groupconsisting

(1) hydrogen,

(2) methyl, and

(3) ethyl.

Another embodiment provides a method for inhibiting phosphorylation ofAkt in a human or animal subject, comprising administering to a human oranimal subject an effective amount of a compound of any one of theembodiments provided herein.

Another embodiment provides a composition, comprising a pharmaceuticallyacceptable carrier and an amount of a compound of any one of theembodiments provided herein effective to inhibit PI3-K activity in ahuman or animal subject when administered thereto. In a more particularembodiment thereof, the composition is effective to inhibit PI3-K alphaactivity in a human or animal subject when administered thereto.

Another embodiment provides a composition, comprising a pharmaceuticallyacceptable carrier, an amount of a compound of any one of theembodiments provided herein effective to inhibit PI3-K activity in ahuman or animal subject when administered thereto, and at least oneadditional agent for the treatment of cancer. In a more particularembodiment thereof, at least one additional agent for the treatment ofcancer is vatalanib (PTK-787), imatinib or gefitinib. Alternatively, theat least one additional agent for the treatment of cancer is selectedfrom the kinase inhibitors, anti-estrogens, anti-androgens, otherinhibitors, cancer chemotherapeutic drugs, alkylating agents, chelatingagents, biological response modifiers, cancer vaccines, or antisensetherapies (groups A-J) listed below. Further, the at least oneadditional agent for the treatment of cancer is selected from radiation,nucleoside analogues, or antimitotic agents.

Another embodiment provides a method for treating a condition bymodulation of PI3-K activity comprising administering to a human oranimal subject in need of such treatment an effective amount of acompound of any one of the embodiments provided herein. In a moreparticular embodiment, the compound has an IC₅₀ value of less than about1 μM with respect to inhibition of PI3K. In another more particularembodiment, the condition is cancer.

Another embodiment provides a method for inhibiting PI3-K activity in ahuman or animal subject, comprising administering to the human or animalsubject a composition comprising an amount of a compound of any one ofthe embodiments provided herein effective to inhibit PI3-K activity thehuman or animal subject.

Another embodiment provides a method for treating a cancer disorder in ahuman or animal subject, comprising administering to the human or animalsubject a composition comprising an amount of a compound of any one ofthe embodiments provided herein effective to inhibit PI3-K activity thehuman or animal subject. A more particular embodiment further comprisesadministering to the human or animal subject at least one additionalagent for the treatment of cancer. In another embodiment, the at leastone additional agent for the treatment of cancer is vatalanib, imatinibor gefitinib. Alternatively, the at least one additional agent for thetreatment of cancer is selected from the kinase inhibitors,anti-estrogens, anti-androgens, other inhibitors, cancerchemotherapeutic drugs, alkylating agents, chelating agents, biologicalresponse modifiers, cancer vaccines, or antisense therapies (groups A-J)listed below.

In another embodiment of any of the aforementioned, the cancer is breastcancer, bladder cancer, colon cancer, glioma, glioblastoma, lung cancer,hepatocellular cancer, gastric cancer, melanoma, thyroid cancer,endometrial cancer, renal cancer, cervical cancer, pancreatic cancer,esophageal cancer, prostate cancer, brain cancer, or ovarian cancer.

Another embodiment provides a method for modulating phosphorylation ofAkt comprising contacting a compound of any one of the embodimentsdescribed herein with a cell. Another embodiment provides a method formodulating phosphorylation of Akt comprising contacting a cell with acompound of any one of the embodiments described herein. In a moreparticular embodiment thereof, said modulation is inhibiting. In a moreparticular embodiment, the compound has an EC₅₀ value of less than about1 μM with respect to inhibition of pAKT. In a more particular embodimentstill, the compound has an EC₅₀ value of less than about 0.5 μM withrespect to inhibition of pAKT. In an even more particular embodiment,the compound has an EC₅₀ value of less than about 0.1 μM with respect toinhibition of pAKT.

Another embodiment provides a compound of any one of the embodimentsdescribed herein for use in the treatment of cancer.

Another embodiment provides for the use of a compound of any one of theembodiments described herein in the manufacture of a medicament for thetreatment for cancer.

Another embodiment provides a method of modulating phosphorylation ofAkt comprising contacting a compound of the present invention with acell. In a more particular embodiment thereof, the compound has an EC₅₀value of less than about 1 μM with respect to inhibition of pAKT.

Another embodiment provides a compound of any one of the embodimentsdescribed herein, and a package insert or other labeling includingdirections for treating a cellular proliferative disease byadministering a PI3-K inhibitory amount of the compound.

The invention further provides compositions, kits, methods of use, andmethods of manufacture as described in the detailed description of theinvention.

BRIEF DESCRIPTION OF THE DRAWINGS

The foregoing aspects and many of the attendant advantages of thisinvention will become more readily appreciated as the same become betterunderstood by reference to the following detailed description, whentaken in conjunction with the accompanying drawings, wherein:

FIG. 1 is a graph illustrating tumor growth inhibition for arepresentative compound of the invention at two dosages compared to acontrol vehicle;

FIG. 2 is a graph illustrating tumor growth inhibition for arepresentative compound of the invention at three dosages compared to acontrol vehicle;

FIG. 3 is a graph illustrating tumor growth inhibition for arepresentative compound of the invention at two dosages compared to acontrol vehicle;

FIG. 4 is a graph illustrating tumor growth inhibition for arepresentative compound of the invention at two dosages compared to acontrol vehicle; and

FIG. 5 is a graph illustrating tumor growth inhibition for arepresentative compound of the invention compared to a control vehicle.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

Phosphotidylinositol-3-kinase (PI3K) mediates the signal from variousgrowth factors to regulate cell proliferation and survival. ASerine/Threonine (Ser/Thr, or S/T) protein kinase, termed Akt, isidentified as a downstream target of PI 3-kinase. This protein kinase isrecruited to the cell membrane by interaction of its pleckstrin homologydomain with PI3K products, phosphatidylinositol-3,4,5-triphosphate(PIP₃), and phosphatidylinositol-3,4-biphosphate (PIP₂), where it isactivated by phosphorylation of its catalytic domain by3-Phosphoinositide-dependent Kinase-1 (PDK-1). Akt is further activatedby phosphorylation of a serine in its C-terminal hydrophobic motif byanother candidate kinase (PDK-2). The activation of Akt acts downstreamto regulate additional kinases many of which are implicated in cellularprocesses that control survival, proliferation, metabolism and growthtranslation. PI3K can also drive cellular processes that impacttransformation, cellular proliferation, cytoskeletal rearrangement andsurvival through a parallel pathway that does not involve Akt (Hennessyet al., Nat. Rev. Drug Disc. 4:988-1004 (2005)). Two of these pathwaysare activation of the small GTP-binding proteins Cdc42 and Rac1 andactivation of the serum and glucocorticoid-inducible kinase (SGK). Cdc42and Rac1, which regulate cytoskeletal movement and cell motility and canfunction as oncogenes when over-expressed, are also linked to the RASpathway. Thus, PI3K activity generates 3′-phosphatidylinositol lipidsthat act as a nodal point to stimulate a diversity of downstreamsignaling pathways.

That these pathways impact cellular properties proliferation, survival,motility and morphology that are often disrupted in cancer,proliferative diseases, thrombotic diseases and inflammation, amongothers, suggests that compounds inhibiting PI3K (and isoforms thereof)have utility, either as a single agent or in combination, in thetreatment of these diseases. In cancer, deregulation of the PI3K/Aktpathway is extensively documented, including overexpression of thePIK3CA gene, activating mutations of the PIK3CA gene, overexpression ofAkt, mutations of PDK-1, and deletions/inactivation of PTEN (Parsons etal., Nature 436:792 (2005); Hennessy et al., Nat. Rev. Drug Disc. 4:988(2005); Stephens et al., Curr. Opin. Pharmacol. 5:1 (2005); Bonneau andLongy, Human Mutation 16:109 (2000) and Ali et al., J. Natl. Can. Inst.91:1922 (1999)). Recent findings indicate that PIK3CA is frequentlymutated (>30%) in various solid tumors in humans (Samuels and Ericson,Curr. Opin. Oncology 18:77 (2005)) and the most frequent of thesemutations promote cell growth and invasion (Samuels et al., Cancer Cell7:561 (2005), and are transforming (Kang et al., Proc. Natl. Acad. Sci.USA 102:802 (2005), Zhao et al., Proc. Natl. Acad. Sci. USA 102:18443(2005)). Thus, inhibitors of PI3K, particularly of the p110α isoformencoded by PIK3CA and its mutations, will be useful in the treatment ofcancers driven by these mutations and deregulations.

The present invention provides novel compounds that act as inhibitors ofserine threonine kinases, lipid kinases, and, more particularly, asinhibitors of phosphatidylinositol 3-kinase (PI3K) function. Thecompounds provided herein can be formulated into pharmaceuticalformulations that are useful in treating patients with a need for aninhibitor of PI3K, especially, in particular embodiments, to providecompositions and methods for reducing cellular proliferation andincreasing cell death in the treatment of cancer.

In one aspect of the present invention, new phosphatidylinositol3-kinase (PI3K) inhibitor compounds, their pharmaceutically acceptablesalts, and prodrugs thereof are provided. The PI3K inhibitor compoundsare pyrimidine-based compounds. The pyrimidine compounds,pharmaceutically acceptable salts, and prodrugs are PI3K inhibitors andare useful in the treating cellular proliferative diseases.

In one embodiment, the phosphatidylinositol 3-kinase (PI3K) inhibitorcompounds of the invention have the formula (I):

or a stereoisomer, tautomer, or pharmaceutically acceptable saltthereof, wherein,

W is CR_(w) or N, wherein R_(w) is selected from the group consisting of

(1) hydrogen,

(2) cyano,

(3) halogen,

(4) methyl,

(5) trifluoromethyl, and

(6) sulfonamido;

R₁ is selected from the group consisting of

(1) hydrogen,

(2) cyano,

(3) nitro,

(4) halogen,

(5) substituted and unsubstituted alkyl,

(6) substituted and unsubstituted alkenyl,

(7) substituted and unsubstituted alkynyl,

(8) substituted and unsubstituted aryl,

(9) substituted and unsubstituted heteroaryl,

(10) substituted and unsubstituted heterocyclyl,

(11) substituted and unsubstituted cycloalkyl,

(12) —COR_(1a),

(13) —CO₂R_(1a),

(14) —CONR_(1a)R_(1b),

(15) —NR_(1a)R_(1b),

(16) —NR_(1a)COR_(1b),

(17) —NR_(1a)SO₂R_(1b),

(18) —OCOR_(1a),

(19) —OR_(1a),

(20) —SR_(1a),

(21) —SOR_(1a),

(22) —SO₂R_(1a), and

(23) —SO₂NR_(1a)R_(1b),

wherein R_(1a), and R_(1b) are independently selected from the groupconsisting of

(a) hydrogen,

(b) substituted or unsubstituted alkyl,

(c) substituted and unsubstituted aryl,

(d) substituted and unsubstituted heteroaryl,

(e) substituted and unsubstituted heterocyclyl, and

(f) substituted and unsubstituted cycloalkyl;

R₂ is selected from the group consisting

(1) hydrogen,

(2) cyano,

(3) nitro,

(4) halogen,

(5) hydroxy,

(6) amino,

(7) substituted and unsubstituted alkyl,

(8) —COR_(2a), and

(9) —NR_(2a)COR_(2b),

wherein R_(2a), and R_(2b) are independently selected from the groupconsisting of

(a) hydrogen, and

(b) substituted or unsubstituted alkyl;

R₃ is selected from the group consisting of

(1) hydrogen,

(2) cyano,

(3) nitro,

(4) halogen,

(5) substituted and unsubstituted alkyl,

(6) substituted and unsubstituted alkenyl,

(7) substituted and unsubstituted alkynyl,

(8) substituted and unsubstituted aryl,

(9) substituted and unsubstituted heteroaryl,

(10) substituted and unsubstituted heterocyclyl,

(11) substituted and unsubstituted cycloalkyl,

(12) —COR_(3a),

(13) —NR_(3a)R_(3b),

(14) —NR_(3a)COR_(3b),

(15) —NR_(3a)SO₂R_(3b),

(16) —OR_(3a),

(17) —SR_(3a),

(18) —SOR_(3a),

(19) —SO₂R_(3a), and

(20) —SO₂NR_(3a)R_(3b),

wherein R_(3a), and R_(3b) are independently selected from the groupconsisting of

(a) hydrogen,

(b) substituted or unsubstituted alkyl,

(c) substituted and unsubstituted aryl,

(d) substituted and unsubstituted heteroaryl,

(e) substituted and unsubstituted heterocyclyl, and

(f) substituted and unsubstituted cycloalkyl; and

R₄ is selected from the group consisting of

(1) hydrogen, and

(2) halogen.

Substituted R₁ comprises substituted or unsubstituted arylalkyl,substituted or unsubstituted heteroarylalkyl, substituted orunsubstituted cycloalkylalkyl, or substituted or unsubstitutedheterocyclylalkyl.

In one embodiment, W is CH.

In another embodiment, W is N. In a more particular embodiment thereof,R₃ is ═O.

In one embodiment, R₁ is selected from the group consisting of

(1) substituted and unsubstituted alkyl,

(2) substituted and unsubstituted aryl,

(3) substituted and unsubstituted heteroaryl,

(4) substituted and unsubstituted heterocyclyl,

(5) substituted and unsubstituted cycloalkyl,

(6) —OR_(1a), and

(7) —NR_(1a)R_(1b),

wherein R_(1a) and R_(1b) are independently selected from the groupconsisting of

(a) substituted and unsubstituted heteroaryl, and

(b) substituted and unsubstituted heterocyclyl.

In another embodiment R₁ is substituted or unsubstituted heterocyclyl,or substituted or unsubstituted —O-heterocyclyl. In another embodiment,R₁ is substituted or unsubstituted morpholinyl; more particular still,R₁ is unsubstituted N-linked morpholinyl.

In another embodiment, R₁ is substituted or unsubstitutedtetrahydropyran or substituted or unsubstituted tetrahydropyranyloxy.More particular still, R₁ is unsubstituted 4-tetrahydropyranyloxy.

In another embodiment, R₁ is substituted or unsubstitutedtetrahydrofuran or substituted or unsubstituted tetrahydrofuranyloxy.More particular still, R₁ is unsubstituted 3-tetrahydrofuranyloxy.

In one embodiment, R₁ comprises substituted or unsubstitutedheterocyclylalkyl, or substituted or unsubstituted heteroarylalkyl. Inone embodiment, R₁ comprises substituted or unsubstituted morpholinyl.In one embodiment, morpholinyl comprises N-linked morpholinyl. In oneembodiment, R₁ comprises substituted or unsubstituted tetrahydropyran.In one embodiment, tetrahydropyran comprises 4-tetrahydropyranyloxy. Inone embodiment, tetrahydropyran comprises 3-tetrahydropyranyloxy. In oneembodiment, R₁ comprises substituted or unsubstituted tetrahydrofuran.In one embodiment, tetrahydrofuran comprises 3-tetrahydrofuranyloxy. Inone embodiment, R₁ comprises substituted or unsubstituted piperidine. Inone embodiment, piperidine comprises 4-piperidinyloxy. In anotherembodiment, piperidine comprises 3-piperidinyloxy. In one embodiment, R₁comprises substituted or unsubstituted pyrrolidine. In one embodiment,pyrrolidine comprises 3-pyrrolidinyloxy.

In one embodiment, R₂ is selected from the group consisting

(1) hydrogen,

(2) cyano,

(3) hydroxy,

(4) halogen,

(5) amino,

(6) methyl, and

(7) trifluoromethyl.

In one embodiment, R₃ is selected from the group consisting of

(1) cyano,

(2) nitro,

(3) halogen,

(4) hydroxy,

(5) amino, and

(6) trifluoromethyl.

In one embodiment, R₃ is trifluoromethyl. In one embodiment, R₃ iscyano.

In one embodiment, the phosphatidylinositol 3-kinase (PI3K) inhibitorcompounds of the invention have the formula (II):

or a stereoisomer, tautomer, or pharmaceutically acceptable saltthereof, wherein,

W is CR_(w) or N, wherein R_(w) is selected from the group consisting of

(1) hydrogen,

(2) cyano,

(3) halogen,

(4) methyl,

(5) trifluoromethyl, and

(6) sulfonamido;

X is O, S, NH, or a direct bond;

R₂ is selected from the group consisting

(1) hydrogen,

(2) cyano,

(3) nitro,

(4) halogen,

(5) hydroxy,

(6) amino,

(7) substituted and unsubstituted alkyl,

(8) —COR_(2a), and

(9) —NR_(2a)COR_(2b),

wherein R_(2a), and R_(2b) are independently selected from the groupconsisting of

(a) hydrogen, and

(b) substituted or unsubstituted alkyl;

R₃ is selected from the group consisting of

(1) hydrogen,

(2) cyano,

(3) nitro,

(4) halogen,

(5) substituted and unsubstituted alkyl,

(6) substituted and unsubstituted alkenyl,

(7) substituted and unsubstituted alkynyl,

(8) substituted and unsubstituted aryl,

(9) substituted and unsubstituted heteroaryl,

(10) substituted and unsubstituted heterocyclyl,

(11) substituted and unsubstituted cycloalkyl,

(12) —COR_(3a),

(13) —NR_(3a)R_(3b),

(14) —NR_(3a)COR_(3b),

(15) —NR_(3a)SO₂R_(3b),

(16) —OR_(3a),

(17) —SR_(3a),

(18) —SOR_(3a),

(19) —SO₂R_(3a), and

(20) —SO₂NR_(3a)R_(3b),

wherein R_(3a), and R_(3b) are independently selected from the groupconsisting of

(a) hydrogen,

(b) substituted or unsubstituted alkyl,

(c) substituted and unsubstituted aryl,

(d) substituted and unsubstituted heteroaryl,

(e) substituted and unsubstituted heterocyclyl, and

(f) substituted and unsubstituted cycloalkyl;

R₄ is selected from the group consisting of

(1) hydrogen, and

(2) halogen; and

R₅ is selected from the group consisting of

(1) substituted and unsubstituted cycloalkyl,

(2) substituted and unsubstituted heterocyclyl,

(3) substituted and unsubstituted aryl, and

(4) substituted and unsubstituted heteroaryl.

In one embodiment, W is CH.

In one embodiment, W is N. In a more particular embodiment thereof, R₃is ═O.

In one embodiment, R₂ is selected from the group consisting of

(1) hydrogen,

(2) cyano,

(3) hydroxy,

(4) amino,

(5) halogen, and

(6) substituted and unsubstituted C₁₋₃ alkyl.

In one embodiment, R₃ is selected from the group consisting of

(1) hydrogen,

(2) cyano,

(3) —SR_(3a)

(4) halogen,

(5) nitro,

(6) substituted and unsubstituted alkyl,

(7) substituted and unsubstituted alkenyl,

(8) substituted and unsubstituted alkynyl,

(9) —OR_(3a),

(10) —NR_(3a)R_(3b),

(11) —COR_(3a), and

(12) —NR_(3a)COR_(3b),

wherein R_(3a), and R_(3b) are independently selected from the groupconsisting of

(a) hydrogen, and

(b) substituted or unsubstituted alkyl.

In one embodiment, R₃ is trifluoromethyl.

In one embodiment, R₅ is selected from the group consisting of

(1) substituted or unsubstituted morpholinyl,

(2) substituted or unsubstituted tetrahydropyranyl, and

(3) substituted or unsubstituted tetrahydrofuranyl.

In a more particular embodiment thereof, R₅ is N-linked morpholinyl;more particular still, X is a direct link. In another more particularembodiment, R₅ is 4-tetrahydropyranyl; more particular still, X is O. Inanother embodiment, R₅ is 3-tetrahydrofuranyl; more particular still, Xis O.

In one embodiment, the phosphatidylinositol 3-kinase (PI3K) inhibitorcompounds of the invention have the formula (III):

or a stereoisomer, tautomer, or pharmaceutically acceptable saltthereof, wherein,

W is CR_(w) or N, wherein R_(w) is selected from the group consisting of

(1) hydrogen,

(2) cyano,

(3) halogen,

(4) methyl,

(5) trifluoromethyl, and

(6) sulfonamido;

R₂ is selected from the group consisting

(1) hydrogen,

(2) cyano,

(3) nitro,

(4) halogen,

(5) hydroxy,

(6) amino,

(7) substituted and unsubstituted alkyl,

(8) —COR_(2a), and

(9) —NR_(2a)COR_(2b),

wherein R_(2a), and R_(2b) are independently selected from the groupconsisting of

(a) hydrogen, and

(b) substituted or unsubstituted alkyl;

R₃ is selected from the group consisting of

(1) hydrogen,

(2) cyano,

(3) nitro,

(4) halogen,

(5) substituted and unsubstituted alkyl,

(6) substituted and unsubstituted alkenyl,

(7) substituted and unsubstituted alkynyl,

(8) substituted and unsubstituted aryl,

(9) substituted and unsubstituted heteroaryl,

(10) substituted and unsubstituted heterocyclyl,

(11) substituted and unsubstituted cycloalkyl,

(12) —COR_(3a),

(13) —NR_(3a)R_(3b),

(14) —NR_(3a)COR_(3b),

(15) —NR_(3a)SO₂R_(3b),

(16) —OR_(3a),

(17) —SR_(3a),

(18) —SOR_(3a),

(19) —SO₂R_(3a), and

(20) —SO₂NR_(3a)R_(3b),

wherein R_(3a), and R_(3b) are independently selected from the groupconsisting of

(a) hydrogen,

(b) substituted or unsubstituted alkyl,

(c) substituted and unsubstituted aryl,

(d) substituted and unsubstituted heteroaryl,

(e) substituted and unsubstituted heterocyclyl, and

(f) substituted and unsubstituted cycloalkyl;

R₄ is selected from the group consisting of

(1) hydrogen, and

(2) halogen; and

R₆ is selected from the group consisting of

(1) hydrogen,

(2) substituted and unsubstituted alkyl, and

(3) substituted and unsubstituted cycloalkyl.

In one embodiment, R₂ is selected from the group consisting

(1) hydrogen,

(2) cyano,

(3) hydroxy,

(4) halogen,

(5) amino,

(6) methyl, and

(7) trifluoromethyl.

In one embodiment, R₃ is selected from the group consisting of

(1) cyano,

(2) nitro,

(3) halogen,

(4) hydroxy,

(5) amino, and

(6) trifluoromethyl.

In one embodiment, R₅ is selected from the group consisting

(1) hydrogen,

(2) methyl, and

(3) ethyl.

It should be understood that the inhibitor compounds according to theinvention may exhibit the phenomenon of tautomerism. As the chemicalstructures within this specification can only represent one of thepossible tautomeric forms, it should be understood that the inventionencompasses any tautomeric form of the drawn structure.

For the compounds of formulas (I)-(III), representative substitutedalkyl groups include arylalkyl, heteroarylalkyl, heterocyclyalkyl,aminoalkyl, alkylaminoalkyl, dialkylaminoalkyl, and sulfonamidoalkylgroups. Representative substituted aryl groups include sulfonamidoarylgroups. Representative substituted heteroaryl groups includealkylheteroaryl groups.

The syntheses of representative PI3K inhibitor compounds of theinvention are described in the methods presented in the Examples Sectionbelow and the preparation of representative compounds are described inExamples 1-31.

Representative PI3K inhibitor compounds of the invention are shown inTable 1.

In other aspects, the present invention provides methods for manufactureof PI3K inhibitor compounds. It is further contemplated that, inaddition to the compounds of formulas (I)-(III), intermediates, andtheir corresponding methods of syntheses are included within the scopeof the invention.

Another embodiment provides a method of inhibiting phosphorylation ofAkt comprising administering a compound of Formula I, II, or III to ahuman in need thereof. Another embodiment provides a method of treatingcancer responsive to inhibition of phosphorylation of Akt, comprisingadministering a compound of Formula I, II, or III. Another embodimentprovides a method of inhibiting phosphorylation of Akt comprisingcontacting a cell with a compound of Formula I, II, or III.

Another embodiment provides a method of inhibiting phosphorylation ofAid comprising orally administering a compound of Formula I, II, or IIIto a human in need thereof. In a more particular embodiment the human issuffering from cancer. In a more particular embodiment the cancer isresponsive to treatment with a compound that inhibits phosphorylation ofAkt. In another embodiment the compound is orally bioavailable.

Another embodiment provides a method of treating cancer comprisingorally administering a compound of Formula I, II, or III, wherein saidcompound is capable of inhibiting activity of pAkt.

In some embodiments of the method of inhibiting PI3K using a PI3Kinhibitor compound of the invention, the IC₅₀ value of the compound isless than or equal to 1 mM with respect to PI3K. In other suchembodiments, the IC₅₀ value is less than or equal to 100 μM, is lessthan or equal to 25 μM, is less than or equal to 10 μM, is less than orequal to 1 μM, is less than or equal to 0.1 μM, is less than or equal to0.050 μM, or is less than or equal to 0.010 μM.

The compounds of the present invention are also useful in assaysevaluating relative activity of PI3 kinase inhibition. In such assays acompound of the present invention can be used to determine relativeinhibitory activity of a compound in comparison to a second compound.When so employed, the compound of the present invention is employed inan amount sufficient to allow the skilled artisan to detect inhibitionof PI3 kinase. Such an amount is sometimes referred to herein as an“effective inhibitory amount.” In a preferred embodiment the inhibitoryamount is an amount that will reduce PI3 kinase activity byapproximately 50% as compared to the activity in the absence of acompound. Other compounds can then be evaluated as providing greater orlesser inhibition at the same concentration so as to provide a rankingof relative activity. Such information is useful in determiningstructural changes and other modifications to the test compound toimprove its activity. Accordingly the present invention provides amethod for inhibiting the activity of PI3 kinase which method comprisescontacting said PI3 kinase with an effective inhibitory amount of acompound of the present invention as disclosed herein. Also provided isa method for inhibiting the activity of PI3 kinase activity in a cell,which method comprises contacting said cell with an effective inhibitoryamount of a compound as claimed herein.

Some embodiments provide methods of inhibiting phosphorylation of Aktusing a compound of the invention having an EC₅₀ value of less thanabout 10 μM with respect to inhibition of pAKT. In another moreparticular embodiment, the compound has an EC₅₀ value of less than about1 μM with respect to inhibition of pAKT. In a more particular embodimentstill, the compound has an EC₅₀ value of less than about 0.5 μM withrespect to inhibition of pAKT. In an even more particular embodiment,the compound has an EC₅₀ value of less than about 0.1 μM with respect toinhibition of pAKT.

In certain embodiments, components of the present invention are capableof inhibition of phosphorylation of Akt. In certain embodiments,components of the invention are capable of inhibition of phosphorylationof Akt in a human or animal subject (i.e., in vivo).

In one embodiment, a method of reducing pAkt activity in a human oranimal subject is provided. In the method, a compound of the inventionis administered in an amount effective to reduce pAkt activity.

In some embodiments of the method of inhibiting PI3K using a PI3Kinhibitor compound of the invention, the EC₅₀ value of the compound isbetween 1 nM to 10 nM. In other such embodiments, the EC₅₀ value isbetween 10 nM to 50 nM, between 50 nM to 100 nM, between 100 nM to 1 μM,between 1 μM to 25 μM, or is between 25 μM to 100 μM.

The compounds of the present invention are also useful in assaysevaluating relative activity of inhibition of phosphorylation of AKT. Insuch assays a compound of the present invention can be used to determinerelative inhibitory activity of a compound in comparison to a secondcompound. When so employed, the compound of the present invention isemployed in an amount sufficient to allow the skilled artisan to detectinhibition AKT phosphorylation. Such an amount is sometimes referred toherein as an “effective inhibitory amount.” In a preferred embodimentthe inhibitory amount is an amount that will reduce phosphorylation ofAKT activity by approximately 50% as compared to the activity in theabsence of a compound. Other compounds can then be evaluated asproviding greater or lesser inhibition at the same concentration so asto provide a ranking of relative activity. Such information is useful indetermining structural changes and other modifications to the testcompound to improve its activity. Accordingly the present inventionprovides a method for inhibiting the AKT phosphorylation which methodcomprises contacting a cell with an effective inhibitory amount of acompound of the present invention, as described herein. Also provided isa method for inhibiting the activity of PI3 kinase activity in a cell,which method comprises contacting said cell with an effective inhibitoryamount of a compound as claimed herein.

In another embodiment, the invention provides methods of treating aPI3K-mediated disorder. In one method, an effective amount of a PI3Kinhibitor compound is administered to a patient (e.g., a human or animalsubject) in need thereof to mediate (or modulate) PI3K activity.

The compounds of the present invention are useful in pharmaceuticalcompositions for human or veterinary, use where inhibition of PI3K isindicated, for example, in the treatment of cellular proliferativediseases such as tumor and/or cancerous cell growth mediated by PI3K. Inparticular, the compounds are useful in the treatment of human or animal(e.g., murine) cancers, including, for example, lung and bronchus;prostate; breast; pancreas; colon and rectum; thyroid; liver andintrahepatic bile duct; hepatocellular; gastric; glioma/glioblastoma;endometrial; melanoma; kidney and renal pelvis; urinary bladder; uterinecorpus; uterine cervix; ovary; multiple myeloma; esophagus; acutemyelogenous leukemia; chronic myelogenous leukemia; lymphocyticleukemia; myeloid leukemia; brain; oral cavity and pharynx; larynx;small intestine; non-Hodgkin lymphoma; melanoma; and villous colonadenoma.

Agents of the invention, in particular, those which have selectivity forPI3 kinase gamma inhibition, are particularly useful in the treatment ofinflammatory or obstructive airways diseases, resulting, for example, inreduction of tissue damage, airways inflammation, bronchialhyperreactivity, remodeling or disease progression. Inflammatory orobstructive airways diseases to which the present invention isapplicable include asthma of whatever type of genesis including bothintrinsic (non-allergic) asthma and extrinsic (allergic) asthma, mildasthma, moderate asthma, severe asthma, bronchitic asthma,exercise-induced asthma, occupational asthma and asthma inducedfollowing bacterial infection. Treatment of asthma is also to beunderstood as embracing treatment of subjects, e.g. of less than 4 or 5years of age, exhibiting wheezing symptoms and diagnosed or diagnosableas “wheezy infants”, an established patient category of major medicalconcern and now often identified as incipient or early-phase asthmatics(“wheezy infant syndrome”).

Compounds of the invention that are selective for one PI3 Kinase isoform(α, β, γ, δ) over a different isoform are compounds that preferentiallyinhibit one isoform. For example, a compound may preferentially inhibitthe alpha isoform over the gamma isoform. Alternatively, a compound maypreferentially inhibit the gamma isoform over the alpha isoform. Todetermine a compound's selectivity, the compound's activity isdetermined according to the Biological Methods described herein. Forexample, the IC₅₀ value, or the EC₅₀ value, of a compound is determinedfor two or more PI3 Kinase isoforms, e.g, alpha and gamma, according toBiological Methods 1 and 2, respectively. The obtained values are thencompared to determine the selectivity of the tested compound.Preferably, the compounds of the invention are selective for one isoformover a second isoform by at least two-fold, five-fold or ten-fold. Evenmore preferably, the compounds of the invention are selective for oneisoform over a second isoform by at least fifty-fold or 100-fold. Evenmore preferably, the compounds of the invention are selective for oneisoform over a second isoform by at least 1000-fold.

Other inflammatory or obstructive airways diseases and conditions towhich the present invention is applicable include acute lung injury(ALI), adult respiratory distress syndrome (ARDS), chronic obstructivepulmonary, airways or lung disease (COPD, COAD or COLD), includingpulmonary fibrosis, chronic bronchitis or dyspnea associated therewith,emphysema, as well as exacerbation of airways hyperreactivity consequentto other drug therapy, in particular other inhaled drug therapy. Theinvention is also applicable to the treatment of bronchitis of whatevertype or genesis including, e.g., acute, arachidic, catarrhal, croupus,chronic or phthinoid bronchitis. Further inflammatory or obstructiveairways diseases to which the present invention is applicable includepneumoconiosis (an inflammatory, commonly occupational, disease of thelungs, frequently accompanied by airways obstruction, whether chronic oracute, and occasioned by repeated inhalation of dusts) of whatever typeor genesis, including, for example, aluminosis, anthracosis, abestosis,chalicosis, ptilosis, siderosis, silicosis, tabacosis and byssinosis.

Having regard to their anti-inflammatory activity, in particular inrelation to inhibition of eosinophil activation, agents of the inventionare also useful in the treatment of eosinophil related disorders, e.g.eosinophilia, in particular eosinophil related disorders of the airways(e.g. involving morbid eosinophilic infiltration of pulmonary tissues)including hypereosinophilia as it effects the airways and/or lungs aswell as, for example, eosinophil-related disorders of the airwaysconsequential or concomitant to Loffler's syndrome, eosinophilicpneumonia, parasitic (in particular metazoan) infestation (includingtropical eosinophilia), bronchopulmonary aspergillosis, polyarteritisnodosa (including Churg-Strauss syndrome), eosinophilic granuloma andeosinophil-related disorders affecting the airways occasioned bydrug-reaction.

Agents of the invention are also useful in the treatment of inflammatoryor allergic conditions of the skin, for example psoriasis, contactdermatitis, atopic dermatitis, alopecia areata, erythema multiforme,dermatitis herpetiformis, scleroderma, vitiligo, hypersensitivityangiitis, urticaria, bullous pemphigoid, lupus erythematosus, pemphisus,epidermolysis bullosa acquisita, and other inflammatory or allergicconditions of the skin.

Agents of the invention may also be used for the treatment of otherdiseases or conditions, in particular diseases or conditions having aninflammatory component, for example, treatment of diseases andconditions of the eye such as conjunctivitis, keratoconjunctivitissicca, and vernal conjunctivitis, diseases affecting the nose includingallergic rhinitis, and inflammatory disease in which autoimmunereactions are implicated or having an autoimmune component or aetiology,including autoimmune haematogical disorders (e.g. haemolytic anaemia,aplastic anaemia, pure red cell anaemia and idiopathicthrombocytopenia), systemic lupus erythematosus, polychondritis,scleroderma, Wegener granulomatosis, dermatomyositis, chronic activehepatitis, myasthenia gravis, Steven-Johnson syndrome, idiopathic sprue,autoimmune inflammatory bowel disease (e.g. ulcerative colitis andCrohn's disease), endocrine ophthalmopathy, Grave's disease,sarcoidosis, alveolitis, chronic hypersensitivity pneumonitis, multiplesclerosis, primary biliary cirrhosis, uveitis (anterior and posterior),interstitial lung fibrosis, psoriatic arthritis and glomerulonephritis(with and without nephritic syndrome, e.g. including idiopathicnephritic syndrome or minal change nephropathy).

In another embodiment, the invention is a method for inhibitingleucocytes, in particular neutrophils and B and T lymphocytes. Exemplarymedical conditions that can be treated include those conditionscharacterized by an undesirable neutrophil function selected from thegroup consisting of stimulated superoxide release, stimulatedexocytosis, and chemotactic migration, preferably without inhibitingphagocytic activity or bacterial killing by the neutrophils.

In another embodiment the invention is a method for disrupting thefunction of osteoclasts and ameliorating a bone resorption disorder,such as osteoporosis.

In another embodiment, diseases or conditions which may be treated withagents of the invention include septic shock, allograft rejectionfollowing transplantation, bone disorders such as but not limited torheumatoid arthritis, ankylosing spondylitis osteoarthritis, obesity,restenosis, diabetes, e.g. diabetes mellitus type I (juvenile diabetes)and diabetes mellitus type II, diarrheal diseases.

In other embodiments, the PI3K-mediated condition or disorder isselected from the group consisting of cardiovascular diseases,atherosclerosis, hypertension, deep venous thrombosis, stroke,myocardial infarction, unstable angina, thromboembolism, pulmonaryembolism, thrombolytic diseases, acute arterial ischemia, peripheralthrombotic occlusions, and coronary artery disease, reperfusioninjuries, retinopathy, such as diabetic retinopathy or hyperbaricoxygen-induced retinopathy, and conditions characterized by elevatedintraocular pressure or secretion of ocular aqueous humor, such asglaucoma.

As described above, since PI3K serves as a second messenger node thatintegrates parallel signaling pathways, evidence is emerging that thecombination of a PI3K inhibitor with inhibitors of other pathways willbe useful in treating cancer and proliferative diseases in humans:

Approximately 20-30% of human breast cancers overexpressHer-2/neu-ErbB2, the target for the drug trastuzumab. Althoughtrastuzumab has demonstrated durable responses in some patientsexpressing Her2/neu-ErbB2, only a subset of these patients respond.Recent work has indicated that this limited response rate can besubstantially improved by the combination of trastuzumab with inhibitorsof PI3K or the PI3K/AKT pathway (Chan et al., Breast Can. Res. Treat.91:187 (2005), Woods Ignatoski et al., Brit. J. Cancer 82:666 (2000),Nagata et al., Cancer Cell 6:117 (2004)).

A variety of human malignancies express activating mutations orincreased levels of Her1/EGFR and a number of antibody and smallmolecule inhibitors have been developed against this receptor tyrosinekinase including tarceva, gefitinib and erbitux. However, while EGFRinhibitors demonstrate anti-tumor activity in certain human tumors(e.g., NSCLC), they fail to increase overall patient survival in allpatients with EGFR-expressing tumors. This may be rationalized by thefact that many downstream targets of Her1/EGFR are mutated orderegulated at high frequencies in a variety of malignancies, includingthe PI3K/Akt pathway. For example, gefitinib inhibits the growth of anadenocarcinoma cell line in in vitro assays. Nonetheless, sub-clones ofthese cell lines can be selected that are resistant to gefitinib thatdemonstrate increased activation of the PI3/Akt pathway. Down-regulationor inhibition of this pathway renders the resistant sub-clones sensitiveto gefitinib (Kokubo et al., Brit. J. Cancer 92:1711 (2005)).Furthermore, in an in vitro model of breast cancer with a cell line thatharbors a PTEN mutation and over-expresses EGFR inhibition of both thePI3K/Akt pathway and EGFR produced a synergistic effect (She et al.,Cancer Cell 8:287-297 (2005)). These results indicate that thecombination of gefitinib and PI3K/Akt pathway inhibitors would be anattractive therapeutic strategy in cancer.

The combination of AEE778 (an inhibitor of Her-2/neu/ErbB2, VEGFR andEGFR) and RAD001 (an inhibitor of mTOR, a downstream target of Akt)produced greater combined efficacy that either agent alone in aglioblastoma xenograft model (Goudar et al., Mol. Cancer. Ther.4:101-112 (2005)).

Anti-estrogens, such as tamoxifen, inhibit breast cancer growth throughinduction of cell cycle arrest that requires the action of the cellcycle inhibitor p27Kip. Recently, it has been shown that activation ofthe Ras-Raf-MAP Kinase pathway alters the phosphorylation status ofp27Kip such that its inhibitory activity in arresting the cell cycle isattenuated, thereby contributing to anti-estrogen resistance (Donovan,et al, J. Biol. Chem. 276:40888, 2001). As reported by Donovan et al.,inhibition of MAPK signaling through treatment with MEK inhibitorreversed the aberrant phosphorylation status of p27 in hormonerefractory breast cancer cell lines and in so doing restored hormonesensitivity. Similarly, phosphorylation of p27Kip by Akt also abrogatesits role to arrest the cell cycle (Viglietto et al., Nat Med. 8:1145(2002)). Accordingly, in one aspect, the compounds of formula (I) may beused in the treatment of hormone dependent cancers, such as breast andprostate cancers, to reverse hormone resistance commonly seen in thesecancers with conventional anticancer agents.

In hematological cancers, such as chronic myelogenous leukemia (CML),chromosomal translocation is responsible for the constitutivelyactivated BCR-Abl tyrosine kinase. The afflicted patients are responsiveto imatinib, a small molecule tyrosine kinase inhibitor, as a result ofinhibition of Abl kinase activity. However, many patients with advancedstage disease respond to imatinib initially, but then relapse later dueto resistance-conferring mutations in the Abl kinase domain. In vitrostudies have demonstrated that BCR-Abl employs the Ras-Raf kinasepathway to elicit its effects. In addition, inhibiting more than onekinase in the same pathway provides additional protection againstresistance-conferring mutations. Accordingly, in another aspect of theinvention, the compounds of formula (I) are used in combination with atleast one additional agent, such as Gleevec®, in the treatment ofhematological cancers, such as chronic myelogenous leukemia (CML), toreverse or prevent resistance to at least one additional agent.

Because activation of the PI3K/Akt pathway drives cell survival,inhibition of the pathway in combination with therapies that driveapoptosis in cancer cells, including radiotherapy and chemotherapy, willresult in improved responses (Ghobrial et al., CA Cancer J. Clin55:178-194 (2005)). As an example, combination of PI3 kinase inhibitorwith carboplatin demonstrated synergistic effects in both in vitroproliferation and apoptosis assays as well as in in vivo tumor efficacyin a xenograft model of ovarian cancer (Westfall and Skinner, Mol.Cancer Ther 4:1764-1771 (2005)).

In addition to cancer and proliferative diseases, there is accumulatingevidence that inhibitors of Class 1A and 1B PI3 kinases would betherapeutically useful in others disease areas. The inhibition of p110γ,the PI3K isoform product of the PIK3CB gene, has been shown to beinvolved in shear-induced platelet activation (Jackson et al., NatureMedicine 11:507-514 (2005)). Thus, a PI3K inhibitor that inhibits p110γwould be useful as a single agent or in combination in anti-thrombotictherapy. The isoform p110γ the product of the PIK3CD gene, is importantin B cell function and differentiation (Clayton et al., J. Exp. Med.196:753-763 (2002)), T-cell dependent and independent antigen responses(Jou et al., Mol. Cell. Biol 22:8580-8590 (2002)) and mast celldifferentiation (Ali et al., Nature 431:1007-1011 (2004)). Thus, it isexpected that p110γ-inhibitors would be useful in the treatment ofB-cell driven autoimmune diseases and asthma. Finally, the inhibition ofp110γ, the isoform product of the PI3KCG gene, results in reduced T, butnot B cell, response (Reif et al., J. Immunol. 173:2236-2240 (2004)) andits inhibition demonstrates efficacy in animal models of autoimmunediseases (Camps et al., Nature Medicine 11:936-943 (2005), Barber etal., Nature Medicine 11:933-935 (2005)).

The present invention provides pharmaceutical compositions comprising atleast one PI3K inhibitor compound (e.g., a compound of formulas(I)-(III)) together with a pharmaceutically acceptable carrier suitablefor administration to a human or animal subject, either alone ortogether with other anticancer agents.

In one embodiment, the present invention provides methods of treatinghuman or animal subjects suffering from a cellular proliferativedisease, such as cancer. The present invention provides methods oftreating a human or animal subject in need of such treatment, comprisingadministering to the subject a therapeutically effective amount of aPI3K inhibitor compound (e.g., a compound of formulas (I)-(III)), eitheralone or in combination with other anticancer agents.

In particular, compositions will either be formulated together as acombination therapeutic or administered separately. Anticancer agentsfor use with the invention include, but are not limited to, one or moreof the following set forth below:

A. Kinase Inhibitors

Kinase inhibitors for use as anticancer agents in conjunction with thecompositions of the present invention include inhibitors of EpidermalGrowth Factor Receptor (EGFR) kinases such as small moleculequinazolines, for example gefitinib (U.S. Pat. No. 5,457,105, U.S. Pat.No. 5,616,582, and U.S. Pat. No. 5,770,599), ZD-6474 (WO 01/32651),erlotinib (Tarceva®, U.S. Pat. No. 5,747,498 and WO 96/30347), andlapatinib (U.S. Pat. No. 6,727,256 and WO 02/02552); VascularEndothelial Growth Factor Receptor (VEGFR) kinase inhibitors, includingSU-11248 (WO 01/60814), SU 5416 (U.S. Pat. No. 5,883,113 and WO99/61422), SU 6668 (U.S. Pat. No. 5,883,113 and WO 99/61422), CHIR-258(U.S. Pat. No. 6,605,617 and U.S. Pat. No. 6,774,237), vatalanib orPTK-787 (U.S. Pat. No. 6,258,812), VEGF-Trap (WO 02/57423),B43-Genistein (WO-09606116), fenretinide (retinoic acidp-hydroxyphenylamine) (U.S. Pat. No. 4,323,581), IM-862 (WO 02/62826),bevacizumab or Avastin® (WO 94/10202), KRN-951,3-[5-(methylsulfonylpiperadine methyl)-indolyl]-quinolone, AG-13736 andAG-13925, pyrrolo[2,1-f][1,2,4]triazines, ZK-304709, Veglin®, VMDA-3601,EG-004, CEP-701 (U.S. Pat. No. 5,621,100), Cand5 (WO 04/09769); Erb2tyrosine kinase inhibitors such as pertuzumab (WO 01/00245),trastuzumab, and rituximab; Akt protein kinase inhibitors, such asRX-0201; Protein Kinase C (PKC) inhibitors, such as LY-317615 (WO95/17182), and perifosine (US 2003171303); Raf/Map/MEK/Ras kinaseinhibitors including sorafenib (BAY 43-9006), ARQ-350RP, LErafAON,BMS-354825 AMG-548, and others disclosed in WO 03/82272; FibroblastGrowth Factor Receptor (FGFR) kinase inhibitors; Cell Dependent Kinase(CDK) inhibitors, including CYC-202 or roscovitine (WO 97/20842 and WO99/02162); Platelet-Derived Growth Factor Receptor (PGFR) kinaseinhibitors such as CHIR-258, 3G3 mAb, AG-13736, SU-11248 and SU6668; andBcr-Abl kinase inhibitors and fusion proteins such as STI-571 orGleevec® (imatinib).

B. Anti-Estrogens

Estrogen-targeting agents for use in anticancer therapy in conjunctionwith the compositions of the present invention include SelectiveEstrogen Receptor Modulators (SERMs) including tamoxifen, toremifene,raloxifene; aromatase inhibitors including Arimidex® or anastrozole;Estrogen Receptor Downregulators (ERDs) including Faslodex® orfulvestrant.

C. Anti-Androgens

Androgen-targeting agents for use in anticancer therapy in conjunctionwith the compositions of the present invention include flutamide,bicalutamide, finasteride, aminoglutethamide, ketoconazole, andcorticosteroids.

D. Other Inhibitors

Other inhibitors for use as anticancer agents in conjunction with thecompositions of the present invention include protein farnesyltransferase inhibitors including tipifarnib or R-115777 (US 2003134846and WO 97/21701), BMS-214662, AZD-3409, and FTI-277; topoisomeraseinhibitors including merbarone and diflomotecan (BN-80915); mitotickinesin spindle protein (KSP) inhibitors including SB-743921 andMKI-833; protease modulators such as bortezomib or Velcade® (U.S. Pat.No. 5,780,454), XL-784; and cyclooxygenase 2 (COX-2) inhibitorsincluding non-steroidal antiinflammatory drugs I (NSAIDs).

E. Cancer Chemotherapeutic Drugs

Particular cancer chemotherapeutic agents for use as anticancer agentsin conjunction with the compositions of the present invention includeanastrozole (Arimidex®), bicalutamide (Casodex®), bleomycin sulfate(Blenoxane®), busulfan (Myleran®), busulfan injection (Busulfex®),capecitabine (Xeloda®), N4-pentoxycarbonyl-5-deoxy-5-fluorocytidine,carboplatin (Paraplatin®), carmustine (BiCNU®), chlorambucil(Leukeran®), cisplatin (Platinol®), cladribine (Leustatin®),cyclophosphamide (Cytoxan® or Neosar®), cytarabine, cytosine arabinoside(Cytosar-U®), cytarabine liposome injection (DepoCyt®), dacarbazine(DTIC-Dome®), dactinomycin (Actinomycin D, Cosmegan), daunorubicinhydrochloride (Cerubidine®), daunorubicin citrate liposome injection(DaunoXome®), dexamethasone, docetaxel (Taxotere®, US 2004073044),doxorubicin hydrochloride (Adriamycin®, Rubex®), etoposide (Vepesid®),fludarabine phosphate (Fludara®), 5-fluorouracil (Adrucil®, Efudex®),flutamide (Eulexin®), tezacitibine, Gemcitabine (difluorodeoxycitidine),hydroxyurea (Hydrea®), Idarubicin (Idamycin®), ifosfamide (IFEX®),irinotecan (Camptosar®), L-asparaginase (ELSPAR®), leucovorin calcium,melphalan (Alkeran®), 6-mercaptopurine (Purinethol®), methotrexate(Folex®), mitoxantrone (Novantrone®), mylotarg, paclitaxel (Taxol®),phoenix (Yttrium90/MX-DTPA), pentostatin, polifeprosan 20 withcarmustine implant (Gliadel®), tamoxifen citrate (Nolvadex®), teniposide(Vumon®), 6-thioguanine, thiotepa, tirapazamine (Tirazone®), topotecanhydrochloride for injection (Hycamptin®), vinblastine (Velban®),vincristine (Oncovin®), and vinorelbine (Navelbine®).

F. Alkylating Agents

Alkylating agents for use in conjunction with the compositions of thepresent invention for anticancer therapeutics include VNP-40101M orcloretizine, oxaliplatin (U.S. Pat. No. 4,169,846, WO 03/24978 and WO03/04505), glufosfamide, mafosfamide, etopophos (U.S. Pat. No.5,041,424), prednimustine; treosulfan; busulfan; irofluven(acylfulvene); penclomedine; pyrazoloacridine (PD-115934);O6-benzylguanine; decitabine (5-aza-2-deoxycytidine); brostallicin;mitomycin C (MitoExtra); TLK-286 (Telcyta®); temozolomide; trabectedin(U.S. Pat. No. 5,478,932); AP-5280 (Platinate formulation of Cisplatin);porfiromycin; and clearazide (meclorethamine).

G. Chelating Agents

Chelating agents for use in conjunction with the compositions of thepresent invention for anticancer therapeutics include tetrathiomolybdate(WO 01/60814); RP-697; Chimeric T84.66 (cT84.66); gadofosveset(Vasovist®); deferoxamine; and bleomycin optionally in combination withelectroporation (EPT).

H. Biological Response Modifiers

Biological response modifiers, such as immune modulators, for use inconjunction with the compositions of the present invention foranticancer therapeutics include staurosprine and macrocyclic analogsthereof; including UCN-01, CEP-701 and midostaurin (see WO 02/30941, WO97/07081, WO 89/07105, U.S. Pat. No. 5,621,100, WO 93/07153, WO01/04125, WO 02/30941, WO 93/08809, WO 94/06799, WO 00/27422, WO96/13506 and WO 88/07045); squalamine (WO 01/79255); DA-9601 (WO98/04541 and U.S. Pat. No. 6,025,387); alemtuzumab; interferons (e.g.IFN-a, IFN-b etc.); interleukins, specifically IL-2 or aldesleukin aswell as IL-1, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11,IL-12, and active biological variants thereof having amino acidsequences greater than 70% of the native human sequence; altretamine(Hexalen®); SU 101 or leflunomide (WO 04/06834 and U.S. Pat. No.6,331,555); imidazoquinolines such as resiquimod and imiquimod (U.S.Pat. Nos. 4,689,338, 5,389,640, 5,268,376, 4,929,624, 5,266,575,5,352,784, 5,494,916, 5,482,936, 5,346,905, 5,395,937, 5,238,944, and5,525,612); and SMIPs, including benzazoles, anthraquinones,thiosemicarbazones, and tryptanthrins (WO 04/87153, WO 04/64759, and WO04/60308).

I. Cancer Vaccines:

Anticancer vaccines for use in conjunction with the compositions of thepresent invention include Avicine® (Tetrahedron Letters 26, 19742269-70); oregovomab (OvaRex®); Theratope® (STn-KLH); Melanoma Vaccines;GI-4000 series (GI-4014, GI-4015, and GI-4016), which are directed tofive mutations in the Ras protein; GlioVax-1; MelaVax; Advexin® orINGN-201 (WO 95/12660); Sig/E7/LAMP-1, encoding HPV-16 E7; MAGE-3Vaccine or M3TK (WO 94/05304); HER-2VAX; ACTIVE, which stimulatesT-cells specific for tumors; GM-CSF cancer vaccine; and Listeriamonocytogenes-based vaccines.

J. Antisense Therapy:

Anticancer agents for use in conjunction with the compositions of thepresent invention also include antisense compositions, such as AEG-35156(GEM-640); AP-12009 and AP-11014 (TGF-beta2-specific antisenseoligonucleotides); AVI-4126; AVI-4557; AVI-4472; oblimersen(Genasense®); JFS2; aprinocarsen (WO 97/29780); GTI-2040 (R2ribonucleotide reductase mRNA antisense oligo) (WO 98/05769); GTI-2501(WO 98/05769); liposome-encapsulated c-Raf antisenseoligodeoxynucleotides (LErafAON) (WO 98/43095); and Sirna-027(RNAi-based therapeutic targeting VEGFR-1 mRNA).

The compounds of the invention can also be combined in a pharmaceuticalcomposition with bronchiodilatory or antihistamine drugs substances.Such bronchiodilatory drugs include anticholinergic or antimuscarinicagents, in particular ipratropium bromide, oxitropium bromide, andtiotropium bromide, and β-2-adrenoreceptor agonists such as salbutamol,terbutaline, salmeterol and, especially, formoterol. Co-therapeuticantihistamine drug substances include cetirizine hydrochloride,clemastine fumarate, promethazine, loratadine, desloratadine,diphenhydramine and fexofenadine hydrochloride.

The effectiveness of an agent of the invention in inhibitinginflammatory conditions, for example in inflammatory airways diseases,may be demonstrated in an animal model, e.g. a mouse or rat model, ofairways inflammation or other inflammatory conditions, for example asdescribed by Szarka et al, J. Immunol. Methods (1997) 202:49-57; Renziet al, Am. Rev. Respir. Dis. (1993) 148:932-939; Tsuyuki et al., J.Clin. Invest. (1995) 96:2924-2931; and Cernadas et al (1999) Am. J.Respir. Cell Mol. Biol. 20:1-8.

The agents of the invention are also useful as co-therapeutic agents foruse in combination with other drug substances such as anti-inflammatory,bronchodilatory or antihistamine drug substances, particularly in thetreatment of obstructive or inflammatory airways diseases such as thosementioned hereinbefore, for example as potentiators of therapeuticactivity of such drugs or as a means of reducing required dosaging orpotential side effects of such drugs. An agent of the invention may bemixed with the other drug substance in a fixed pharmaceuticalcomposition or it may be administered separately, before, simultaneouslywith or after the other drug substance. Accordingly the inventionincludes a combination of an agent of the invention as hereinbeforedescribed with an anti-inflammatory, bronchodilatory or antihistaminedrug substance, said agent of the invention and said drug substancebeing in the same or different pharmaceutical composition. Suchanti-inflammatory drugs include steroids, in particularglucocorticosteroids such as budesonide, beclamethasone, fluticasone,ciclesonide or mometasone, LTB4 antagonists such as those described inU.S. Pat. No. 5,451,700, LTD4 antagonists such as montelukast andzafirlukast, dopamine receptor agonists such as cabergoline,bromocriptine, ropinirole and4-hydroxy-7-[2-[[2-[[3-(2-phenylethoxy)propyl]-sulfonyl]ethyl]-amino]ethyl]-2(3H)-benzothiazoloneand pharmaceutically acceptable salts thereof (the hydrochloride beingViozan®—AstraZeneca), and PDE4 inhibitors such as Ariflo® (GlaxoSmithKline), Roflumilast (Byk Gulden), V-11294A (Napp), BAY19-8004 (Bayer),SCH-351591 (Schering-Plough), Arofylline (Almirall Prodesfarma) andPD189659 (Parke-Davis). Such bronchodilatory drugs includeanticholinergic or antimuscarinic agents, in particular ipratropiumbromide, oxitropium bromide and tiotropium bromide, and beta-2adrenoceptor agonists such as salbutamol, terbutaline, salmeterol and,especially, formoterol and pharmaceutically acceptable salts thereof,and compounds (in free or salt or solvate form) of formula I of PCTInternational patent publication No. WO 00/75114, which document isincorporated herein by reference, preferably compounds of the Examplesthereof, especially a compound of formula

and pharmaceutically acceptable salts thereof. Co-therapeuticantihistamine drug substances include cetirizine hydrochloride,acetaminophen, clemastine fumarate, promethazine, loratidine,desloratidine, diphenhydramine and fexofenadine hydrochloride.Combinations of agents of the invention and steroids, beta-2 agonists,PDE4 inhibitors or LTD4 antagonists may be used, for example, in thetreatment of COPD or, particularly, asthma. Combinations of agents ofthe invention and anticholinergic or antimuscarinic agents, PDE4inhibitors, dopamine receptor agonists or LTB4 antagonists may be used,for example, in the treatment of asthma or, particularly, COPD.

Other useful combinations of agents of the invention withanti-inflammatory drugs are those with antagonists of chemokinereceptors, e.g. CCR-1, CCR-2, CCR-3, CCR-4, CCR-5, CCR-6, CCR-7, CCR-8,CCR-9 and CCR10, CXCR1, CXCR2, CXCR3, CXCR4, CXCR5, particularly CCR-5antagonists such as Schering-Plough antagonists SC-351125, SCH-55700 andSCH-D, Takeda antagonists such asN-[[4-[[[6,7-dihydro-2-(4-methylphenyl)-5H-benzocyclohepten-8-yl]carbonyl]amino]phenyl]-methyl]tetrahydro-N,N-dimethyl-2H-pyran-4-aminiumchloride (TAK-770), and CCR-5 antagonists described in U.S. Pat. No.6,166,037 (particularly claims 18 and 19), WO 00/66558 (particularlyclaim 8), and WO 00/66559 (particularly claim 9).

The compounds of the invention can also be combined in a pharmaceuticalcomposition with compounds that are useful for the treatment of athrombolytic disease, heart disease, stroke, etc., (e.g., aspirin,streptokinase, tissue plasminogen activator, urokinase, anticoagulants,antiplatelet drugs (e.g, PLAVIX; clopidogrel bisulfate), a statin (e.g.,LIPITOR or Atorvastatin calcium), ZOCOR (Simvastatin), CRESTOR(Rosuvastatin), etc.), a Beta blocker (e.g., Atenolol), NORVASC(amlodipine besylate), and an ACE inhibitor (e.g., lisinopril).

The compounds of the invention can also be combined in a pharmaceuticalcomposition with compounds that are useful for the treatment ofantihypertension agents such as, ACE inhibitors, lipid lowering agentssuch as statins, LIPITOR (Atorvastatin calcium), calcium channelblockers dush as NORVASC (amlodipine besylate). The compounds of thepresent invention may also be used in combination with fibrates,beta-blockers, NEPI inhibitors, Angiotensin-2 receptor antagonists andplatelet aggregation inhibitors.

For the treatment of inflammatory diseases, including rheumatoidarthritis, the compounds of the invention may be combined with agentssuch as TNF-α inhibitors such as anti-TNF-α monoclonal antibodies (suchas REMICADE, CDP-870) and D2E7 (HUMIRA) and TNF receptor immunoglobulinfusion molecules (such as ENBREL), IL-1 inhibitors, receptor antagonistsor soluble IL-1Rα (e.g. KINERET or ICE inhibitors), nonsterodialanti-inflammatory agents (NSAIDS), piroxicam, diclofenac, naproxen,flurbiprofen, fenoprofen, ketoprofen ibuprofen, fenamates, mefenamicacid, indomethacin, sulindac, apazone, pyrazolones, phenylbutazone,aspirin, COX-2 inhibitors (such as CELEBREX (celecoxib), PREXIGE(lumiracoxib)), metalloprotease inhibitors (preferably MMP-13 selectiveinhibitors), p2X7 inhibitors, α2δ inhibitors, NEUROTIN, pregabalin, lowdose methotrexate, leflunomide, hydroxyxchloroquine, d-penicillamine,auranofin or parenteral or oral gold.

The compounds of the invention can also be used in combination with theexisting therapeutic agents for the treatment of osteoarthritis.Suitable agents to be used in combination include standard non-steroidalanti-inflammatory agents (hereinafter NSAID's) such as piroxicam,diclofenac, propionic acids such as naproxen, flurbiprofen, fenoprofen,ketoprofen and ibuprofen, fenamates such as mefenamic acid,indomethacin, sulindac, apazone, pyrazolones such as phenylbutazone,salicylates such as aspirin, COX-2 inhibitors such as celecoxib,valdecoxib, lumiracoxib and etoricoxib, analgesics and intraarticulartherapies such as corticosteroids and hyaluronic acids such as hyalganand synvisc.

The compounds of the invention may also be used in combination withantiviral agents such as Viracept, AZT, acyclovir and famciclovir, andantisepsis compounds such as Valant.

The compounds of the present invention may also be used in combinationwith CNS agents such as antidepressants (sertraline), anti-Parkinsoniandrugs (such as deprenyl, L-dopa, Requip, Mirapex, MAOB inhibitors suchas selegine and rasagiline, comP inhibitors, such as Tasmar,A-2inhibitors, dopamine reuptake inhibitors, NMDA antagonists, Nicotineagonists, Dopamine agonists, and inhibitors of neuronal nitric oxidesynthase), and anti-Alzheimer's drugs such as donepezil, tacrine, α2δinhibitors, NEUROTIN, pregabalin, COX-2 inhibitors, propentofylline ormetryfonate.

The compounds of the present invention may also be used in combinationwith osteoporosis agents such as EVISTA (raloxifene hydrochloride),droloxifene, lasofoxifene or fosomax and immunosuppressant agents suchas FK-506 and rapamycin.

In another aspect of the invention, kits that include one or morecompounds of the invention are provided. Representative kits include aPI3K inhibitor compound of the invention (e.g., a compound of formulas(I)-(III)) and a package insert or other labeling including directionsfor treating a cellular proliferative disease by administering an PI3Kinhibitory amount of the compound.

The following definitions are provided to better understand theinvention.

“Alkyl” refers to alkyl groups that do not contain heteroatoms. Thus thephrase includes straight chain alkyl groups such as methyl, ethyl,propyl, butyl, pentyl, hexyl, heptyl, octyl, nonyl, decyl, undecyl,dodecyl and the like. The phrase also includes branched chain isomers ofstraight chain alkyl groups, including but not limited to, the followingwhich are provided by way of example: —CH(CH₃)₂, —CH(CH₃)(CH₂CH₃),—CH(CH₂CH₃)₂, —C(CH₃)₃, —C(CH₂CH₃)₃, —CH₂CH(CH₃)₂, —CH₂CH(CH₃)(CH₂CH₃),—CH₂CH(CH₂CH₃)₂, —CH₂C(CH₃)₃, —CH₂C(CH₂CH₃)₃, —CH(CH₃)—CH(CH₃)(CH₂CH₃),—CH₂CH₂CH(CH₃)₂, —CH₂CH₂CH(CH₃)(CH₂CH₃), —CH₂CH₂CH(CH₂CH₃)₂,—CH₂CH₂C(CH₃)₃, —CH₂CH₂C(CH₂CH₃)₃, —CH(CH₃)CH₂₋CH(CH₃)₂,—CH(CH₃)CH(CH₃)CH(CH₃)₂, —CH(CH₂CH₃)CH(CH₃)CH(CH₃)(CH₂CH₃), and others.Thus the phrase “alkyl groups” includes primary alkyl groups, secondaryalkyl groups, and tertiary alkyl groups. Preferred alkyl groups includestraight and branched chain alkyl groups having 1 to 12 carbon atoms or1 to 6 carbon atoms.

“Alkylene” refers to the same residues as noted above for “alkyl,” buthaving two points of attachment. Exemplary alkylene groups includeethylene (—CH₂CH₂—), propylene (—CH₂CH₂CH₂—), dimethylpropylene(—CH₂C(CH₃)₂CH₂—), and cyclohexylpropylene (—CH₂CH₂CH(C₆H₁₃)—).

“Alkenyl” refers to straight chain, branched, or cyclic groups from 2 toabout 20 carbon atoms such as those described with respect to alkylgroups as defined above, except having one or more carbon-carbon doublebonds. Examples include, but are not limited to vinyl, —CH═C(H)(CH₃),—CH═C(CH₃)₂, —C(CH₃)═C(H)₂, —C(CH₃)═C(H)(CH₃), —C(CH₂CH₃)═CH₂,cyclohexenyl, cyclopentenyl, cyclohexadienyl, butadienyl, pentadienyl,and hexadienyl among others. Preferred alkenyl groups include straightchain and branched alkenyl groups and cyclic alkenyl groups having 2 to12 carbon atoms or 2 to 6 carbon atoms.

“Alkynyl” refers to straight chain, branched, or cyclic groups from 2 toabout 20 carbon atoms such as those described with respect to alkylgroups as defined above, except having one or more carbon-carbon triplebonds. Examples include, but are not limited to —C≡C(H), —C≡C(CH₃),—C≡C(CH₂CH₃), —C(H₂)C≡C(H), —C(H)₂C≡C(CH₃), and —C(H)₂C≡C(CH₂CH₃) amongothers. Preferred alkynyl groups include straight chain and branchedalkynyl groups having 2 to 12 carbon atoms or 2 to 6 carbon atoms.

Alkyl, alkenyl, and alkynyl groups may be substituted. “Substitutedalkyl” refers to an alkyl group as defined above in which one or morebonds to a carbon(s) or hydrogen(s) are replaced by a bond tonon-hydrogen and non-carbon atoms such as, but not limited to, a halogenatom such as F, Cl, Br, and I; an oxygen atom in groups such as hydroxylgroups, alkoxy groups, aryloxy groups, and ester groups; a sulfur atomin groups such as thiol groups, alkyl and aryl sulfide groups, sulfonegroups, sulfonyl groups, and sulfoxide groups; a nitrogen atom in groupssuch as amines, amides, alkylamines, dialkylamines, arylamines,alkylarylamines, diarylamines, N-oxides, imides, and enamines; a siliconatom in groups such as in trialkylsilyl groups, dialkylarylsilyl groups,alkyldiarylsilyl groups, and triarylsilyl groups; and other heteroatomsin various other groups. Substituted alkyl groups also include groups inwhich one or more bonds to a carbon(s) or hydrogen(s) atom is replacedby a higher-order bond (e.g., a double- or triple-bond) to a heteroatomsuch as oxygen in oxo, carbonyl, carboxyl, and ester groups; nitrogen ingroups such as imines, oximes, hydrazones, and nitriles. Substitutedalkyl groups further include alkyl groups in which one or more bonds toa carbon(s) or hydrogen(s) atoms is replaced by a bond to an aryl,heteroaryl, heterocyclyl, or cycloalkyl group. Preferred substitutedalkyl groups include, among others, alkyl groups in which one or morebonds to a carbon or hydrogen atom is/are replaced by one or more bondsto fluoro, chloro, or bromo group. Another preferred substituted alkylgroup is the trifluoromethyl group and other alkyl groups that containthe trifluoromethyl group. Other preferred substituted alkyl groupsinclude those in which one or more bonds to a carbon or hydrogen atom isreplaced by a bond to an oxygen atom such that the substituted alkylgroup contains a hydroxyl, alkoxy, or aryloxy group. Other preferredsubstituted alkyl groups include alkyl groups that have an amine, or asubstituted or unsubstituted alkylamine, dialkylamine, arylamine,(alkyl)(aryl)amine, diarylamine, heterocyclylamine, diheterocyclylamine,(alkyl)(heterocyclyl)amine, or (aryl)(heterocyclyl)amine group. Stillother preferred substituted alkyl groups include those in which one ormore bonds to a carbon(s) or hydrogen(s) atoms is replaced by a bond toan aryl, heteroaryl, heterocyclyl, or cycloalkyl group. Examples ofsubstituted alkyl are: —(CH₂)₃NH₂, —(CH₂)₃NH(CH₃), —(CH₂)₃NH(CH₃)₂,—CH₂C(═CH₂)CH₂NH₂, —CH₂C(═O)CH₂NH₂, —CH₂S(═O)₂CH₃, —CH₂OCH₂NH₂, —CO₂H.Examples of substituents of substituted alkyl are: —CH₃, —C₂H₅, —CH₂OH,—OH, —OCH₃, —OC₂H₅, —OCF₃, —OC(═O)CH₃, —OC(═O)NH₂, —OC(═O)N(CH₃)₂, —CN,—NO₂, —C(═O)CH₃, —CO₂H, —CO₂CH₃, —CONH₂, —NH₂, —N(CH₃)₂, —NHSO₂CH₃,—NHCOCH₃, —NHCOCH₃, —NHC(═O)OCH₃, —NHSO—₂CH₃, —SO₂CH₃, —SO₂NH₂, Halo.

“Substituted alkenyl” has the same meaning with respect to alkenylgroups that substituted alkyl groups had with respect to unsubstitutedalkyl groups. A substituted alkenyl group includes alkenyl groups inwhich a non-carbon or non-hydrogen atom is bonded to a carbon doublebonded to another carbon and those in which one of the non-carbon ornon-hydrogen atoms is bonded to a carbon not involved in a double bondto another carbon.

“Substituted alkynyl” has the same meaning with respect to alkynylgroups that substituted alkyl groups had with respect to unsubstitutedalkyl groups. A substituted alkynyl group includes alkynyl groups inwhich a non-carbon or non-hydrogen atom is bonded to a carbon triplebonded to another carbon and those in which a non-carbon or non-hydrogenatom is bonded to a carbon not involved in a triple bond to anothercarbon.

“Alkoxy” refers to RO— wherein R is alkyl. Representative examples ofalkoxy groups include methoxy, ethoxy, t-butoxy, trifluoromethoxy, andthe like.

“Halogen” or “halo” refers to chloro, bromo, fluoro, and iodo groups.The term “haloalkyl” refers to an alkyl radical substituted with one ormore halogen atoms. The term “haloalkoxy” refers to an alkoxy radicalsubstituted with one or more halogen atoms.

“Amino” refers herein to the group —NH₂. The term “alkylamino” refersherein to the group —NRR′ where R is alkyl and R′ is hydrogen or alkyl.The term “arylamino” refers herein to the group —NRR′ where R is aryland R′ is hydrogen, alkyl, or aryl. The term “aralkylamino” refersherein to the group —NRR′ where R is aralkyl and R′ is hydrogen, alkyl,aryl, or aralkyl.

“Alkoxyalkyl” refers to the group -alk₁-O-alk₂ where alk₁ is alkyl oralkenyl, and alk₂ is alkyl or alkenyl. The term “aryloxyalkyl” refers tothe group -alkyl O-aryl. The term “aralkoxyalkyl” refers to the group-alkylenyl-O-aralkyl.

“Alkoxyalkylamino” refers herein to the group —NR-(alkoxyalkyl), where Ris typically hydrogen, aralkyl, or alkyl.

“Aminocarbonyl” refers herein to the group —C(O)—NH₂. “Substitutedaminocarbonyl” refers herein to the group —C(O)—NRR′ where R is alkyland R′ is hydrogen or alkyl. The term “arylaminocarbonyl” refers hereinto the group —C(O)—NRR′ where R is aryl and R′ is hydrogen, alkyl oraryl. “Aralkylaminocarbonyl” refers herein to the group —C(O)—NRR′ whereR is aralkyl and R′ is hydrogen, alkyl, aryl, or aralkyl.

“Aminosulfonyl” refers herein to the group —S(O)₂—NH₂. “Substitutedaminosulfonyl” refers herein to the group —S(O)₂—NRR′ where R is alkyland R′ is hydrogen or alkyl. The term “aralkylaminosulfonlyaryl” refersherein to the group -aryl-S(O)₂—NH-aralkyl.

“Carbonyl” refers to the divalent group —C(O)—.

“Carbonyloxy” refers generally to the group —C(O)—O. Such groups includeesters, —C(O)—O—R, where R is alkyl, cycloalkyl, aryl, or aralkyl. Theterm “carbonyloxycycloalkyl” refers generally herein to both a“carbonyloxycarbocycloalkyl” and a “carbonyloxyheterocycloalkyl,” i.e.,where R is a carbocycloalkyl or heterocycloalkyl, respectively. The term“arylcarbonyloxy” refers herein to the group —C(O)—O-aryl, where aryl isa mono- or polycyclic, carbocycloaryl or heterocycloaryl. The term“aralkylcarbonyloxy” refers herein to the group —C(O)—O-aralkyl.

“Sulfonyl” refers herein to the group —SO₂—. “Alkylsulfonyl” refers to asubstituted sulfonyl of the structure —SO₂R— in which R is alkyl.Alkylsulfonyl groups employed in compounds of the present invention aretypically alkylsulfonyl groups having from 1 to 6 carbon atoms in itsbackbone structure. Thus, typical alkylsulfonyl groups employed incompounds of the present invention include, for example, methylsulfonyl(i.e., where R is methyl), ethylsulfonyl (i.e., where R is ethyl),propylsulfonyl (i.e., where R is propyl), and the like. The term“arylsulfonyl” refers herein to the group —SO₂-aryl. The term“aralkylsulfonyl” refers herein to the group —SO₂-aralkyl. The term“sulfonamido” refers herein to —SO₂NH₂.

“Carbonylamino” refers to the divalent group —NH—C(O)— in which thehydrogen atom of the amide nitrogen of the carbonylamino group can bereplaced alkyl, aryl, or aralkyl group. Such groups include moietiessuch as carbamate esters (—NH—C(O)—O—R) and amides —NH—C(O)—R, where Ris a straight or branched chain alkyl, cycloalkyl, or aryl or aralkyl.The term “alkylcarbonylamino” refers to alkylcarbonylamino where R isalkyl having from 1 to about 6 carbon atoms in its backbone structure.The term “arylcarbonylamino” refers to group —NH—C(O)—R where R is anaryl. Similarly, the term “aralkylcarbonylamino” refers to carbonylaminowhere R is aralkyl.

“Guanidino” or “guanidyl” refers to moieties derived from guanidine,H₂N—C(═NH)—NH₂. Such moieties include those bonded at the nitrogen atomcarrying the formal double bond (the “2”-position of the guanidine,e.g., diaminomethyleneamino, (H₂N)₂C═NH—)) and those bonded at either ofthe nitrogen atoms carrying a formal single bond (the “1-” and/or“3”-positions of the guanidine, e.g., H₂N—C(═NH)—NH—)). The hydrogenatoms at any of the nitrogens can be replaced with a suitablesubstituent, such as alkyl, aryl, or aralkyl.

“Amidino” refers to the moieties R—C(═N)—NR′— (the radical being at the“N¹” nitrogen) and R(NR′)C═N— (the radical being at the “N²” nitrogen),where R and R′ can be hydrogen, alkyl, aryl, or aralkyl.

“Cycloalkyl” refers to a mono- or polycyclic, heterocyclic orcarbocyclic alkyl substituent. Representative cycloalkyl groups includecyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, andcyclooctyl and such rings substituted with straight and branched chainalkyl groups as defined above. Typical cycloalkyl substituents have from3 to 8 backbone (i.e., ring) atoms in which each backbone atom is eithercarbon or a heteroatom. The term “heterocycloalkyl” refers herein tocycloalkyl substituents that have from 1 to 5, and more typically from 1to 4 heteroatoms in the ring structure. Suitable heteroatoms employed incompounds of the present invention are nitrogen, oxygen, and sulfur.Representative heterocycloalkyl moieties include, for example,morpholino, piperazinyl, piperadinyl, and the like. Carbocycloalkylgroups are cycloalkyl groups in which all ring atoms are carbon. Whenused in connection with cycloalkyl substituents, the term “polycyclic”refers herein to fused and non-fused alkyl cyclic structures.

“Substituted heterocycle,” “heterocyclic group,” “heterocycle,” or“heterocyclyl,” as used herein refers to any 3- or 4-membered ringcontaining a heteroatom selected from nitrogen, oxygen, and sulfur or a5- or 6-membered ring containing from one to three heteroatoms selectedfrom the group consisting of nitrogen, oxygen, or sulfur; wherein the5-membered ring has 0-2 double bonds and the 6-membered ring has 0-3double bonds; wherein the nitrogen and sulfur atom maybe optionallyoxidized; wherein the nitrogen and sulfur heteroatoms maybe optionallyquarternized; and including any bicyclic group in which any of the aboveheterocyclic rings is fused to a benzene ring or another 5- or6-membered heterocyclic ring independently defined above. Examples ofheterocyclyl groups include, but are not limited to: unsaturated 3- to8-membered rings containing 1 to 4 nitrogen atoms such as, but notlimited to pyrrolyl, dihydropyridyl, pyrimidyl, pyrazinyl, tetrazolyl,(e.g., 1H-tetrazolyl, 2H-tetrazolyl); condensed unsaturated heterocyclicgroups containing 1 to 4 nitrogen atoms such as, but not limited to,isoindolyl, indolinyl, indolizinyl, quinolyl, indazolyl; unsaturated 3-to 8-membered rings containing 1 to 2 oxygen atoms and 1 to 3 nitrogenatoms such as, but not limited to, oxadiazolyl (e.g., 1,2,4-oxadiazolyl,1,3,4-oxadiazolyl, 1,2,5-oxadiazolyl); saturated 3- to 8-membered ringscontaining 1 to 2 oxygen atoms and 1 to 3 nitrogen atoms such as, butnot limited to, morpholinyl; unsaturated condensed heterocyclic groupscontaining 1 to 2 oxygen atoms and 1 to 3 nitrogen atoms, for example,benzoxadiazolyl, benzoxazinyl (e.g., 2H-1,4-benzoxazinyl); unsaturated3- to 8-membered rings containing 1 to 3 sulfur atoms and 1 to 3nitrogen atoms such as, but not limited to, thiadiazolyl (e.g.,1,2,3-thiadiazolyl, 1,2,4-thiadiazolyl, 1,3,4-thiadiazolyl,1,2,-thiadiazolyl); saturated 3- to 8-membered rings containing 1 to 2sulfur atoms and 1 to 3 nitrogen atoms such as, but not limited to,thiazolodinyl; saturated and unsaturated 3- to 8-membered ringscontaining 1 to 2 sulfur atoms such as, but not limited to,dihydrodithienyl, dihydrodithionyl, tetrahydrothiophene,tetra-hydrothiopyran; unsaturated condensed heterocyclic ringscontaining 1 to 2 sulfur atoms and 1 to 3 nitrogen atoms such as, butnot limited to, benzothiadiazolyl, benzothiazinyl (e.g.,2H-1,4-benzothiazinyl), dihydrobenzothiazinyl (e.g.,2H-3,4-dihydrobenzothiazinyl), unsaturated 3- to 8-membered ringscontaining oxygen atoms such as, but not limited to furyl; unsaturatedcondensed heterocyclic rings containing 1 to 2 oxygen atoms such asbenzodioxoyl (e.g., 1,3-benzodioxoyl); unsaturated 3- to 8-memberedrings containing an oxygen atom and 1 to 2 sulfur atoms such as, but notlimited to, dihydrooxathienyl; saturated 3- to 8-membered ringscontaining 1 to 2 oxygen atoms and 1 to 2 sulfur atoms such as1,4-oxathiane; unsaturated condensed rings containing 1 to 2 sulfuratoms such as benzodithienyl; and unsaturated condensed heterocyclicrings containing an oxygen atom and 1 to 2 oxygen atoms such asbenzoxathienyl. Preferred heterocycles include, for example: diazapinyl,pyrryl, pyrrolinyl, pyrrolidinyl, pyrazolyl, pyrazolinyl, pyrazolidinyl,imidazoyl, imidazolinyl, imidazolidinyl, pyridyl, piperidinyl,pyrazinyl, piperazinyl, N-methyl piperazinyl, azetidinyl,N-methylazetidinyl, pyrimidinyl, pyridazinyl, oxazolyl, oxazolidinyl,isoxazolyl, isoxazolidinyl, morpholinyl, thiazolyl, thiazolidinyl,isothiazolyl, isothiazolidinyl, indolyl, quinolinyl, isoquinolinyl,benzimidazolyl, benzothiazolyl, benzoxazolyl, furyl, thienyl, triazolyl,and benzothienyl. Heterocyclyl groups also include those described abovein which one or more S atoms in the ring is double-bonded to one or twooxygen atoms (sulfoxides and sulfones). For example, heterocyclyl groupsinclude tetrahydrothiophene, tetrahydrothiophene oxide, andtetrahydrothiophene 1,1-dioxide. Preferred heterocyclyl groups contain 5or 6 ring members. More preferred heterocyclyl groups includepiperazine, 1,2,3-triazole, 1,2,4-triazole, tetrazole, thiomorpholine,homopiperazine, oxazolidin-2-one, pyrrolidin-2-one, quinuclidine, andtetrahydrofuran.

Heterocyclic moieties can be unsubstituted or monosubstituted ordisubstituted with various substituents independently selected fromhydroxy, halo, oxo (C═O), alkylimino (RN═, wherein R is alkyl or alkoxygroup), amino, alkylamino, dialkylamino, acylaminoalkyl, alkoxy,thioalkoxy, polyalkoxy, alkyl, cycloalkyl or haloalkyl. “Unsubstitutedheterocyclyl” includes condensed heterocyclic rings such asbenzimidazolyl, it does not include heterocyclyl groups that have othergroups such as alkyl or halo groups bonded to one of the ring members ascompounds such as 2-methylbenzimidazolyl are substituted heterocyclylgroups.

The heterocyclic groups may be attached at various positions as will beapparent to those having skill in the organic and medicinal chemistryarts in conjunction with the disclosure herein.

where R is H or a heterocyclic substituent, as described herein.

Representative heterocyclics include, for example, imidazolyl, pyridyl,piperazinyl, azetidinyl, thiazolyl, furanyl, triazolyl benzimidazolyl,benzothiazolyl, benzoxazolyl, quinolinyl, isoquinolinyl, quinazolinyl,quinoxalinyl, phthalazinyl, indolyl, naphthpyridinyl, indazolyl, andquinolizinyl.

“Aryl” refers to optionally substituted monocyclic and polycyclicaromatic groups having from 3 to 14 backbone carbon or hetero atoms, andincludes both carbocyclic aryl groups and heterocyclic aryl groups. Theterm refers to, but is not limited to, groups such as phenyl, biphenyl,anthracenyl, naphthenyl by way of example. Carbocyclic aryl groups arearyl groups in which all ring atoms in the aromatic ring are carbon. Theterm “heteroaryl” refers herein to aryl groups having from 1 to 4heteroatoms as ring atoms in an aromatic ring with the remainder of thering atoms being carbon atoms.

“Unsubstituted aryl” includes groups containing condensed rings such asnaphthalene. It does not include aryl groups that have other groups suchas alkyl or halo groups bonded to one of the ring members, as arylgroups such as tolyl are considered herein to be substituted aryl groupsas described below. A preferred unsubstituted aryl group is phenyl.Unsubstituted aryl groups may be bonded to one or more carbon atom(s),oxygen atom(s), nitrogen atom(s), and/or sulfur atom(s) in the parentcompound, however.

“Substituted aryl group” has the same meaning with respect tounsubstituted aryl groups that substituted alkyl groups had with respectto unsubstituted alkyl groups. However, a substituted aryl group alsoincludes aryl groups in which one of the aromatic carbons is bonded toone of the non-carbon or non-hydrogen atoms described above and alsoincludes aryl groups in which one or more aromatic carbons of the arylgroup is bonded to a substituted and/or unsubstituted alkyl, alkenyl, oralkynyl group as defined herein. This includes bonding arrangements inwhich two carbon atoms of an aryl group are bonded to two atoms of analkyl, alkenyl, or alkynyl group to define a fused ring system (e.g.,dihydronaphthyl or tetrahydronaphthyl). Thus, the phrase “substitutedaryl” includes, but is not limited to tolyl, and hydroxyphenyl amongothers.

“Substituted heteroaryl” as used herein refers to a heteroaryl group asdefined herein substituted by independent replacement of one, two orthree of the hydrogen atoms thereon with Cl, Br, F, I, —OH, —CN,C₁-C₃-alkyl, C₁-C₆-alkoxy, C₁-C₆-alkoxy substituted with aryl,haloalkyl, thioalkoxy, amino, alkylamino, dialkylamino, mercapto, nitro,carboxaldehyde, carboxy, alkoxycarbonyl and carboxamide. In addition,any one substituent may be an aryl, heteroaryl, or heterocycloalkylgroup.

When used in connection with aryl substituents, the term “polycyclicaryl” refers herein to fused and non-fused cyclic structures in which atleast one cyclic structure is aromatic, such as, for example,benzodioxole (which has a heterocyclic structure fused to a phenylgroup, i.e.,

naphthyl, and the like. Exemplary aryl or heteroaryl moieties employedas substituents in compounds of the present invention include phenyl,pyridyl, pyrimidinyl, thiazolyl, indolyl, imidazolyl, oxadiazolyl,tetrazolyl, pyrazinyl, triazolyl, thiophenyl, furanyl, quinolinyl,purinyl, naphthyl, benzothiazolyl, benzopyridyl, and benzimidazolyl, andthe like.

“Aralkyl” or “arylalkyl” refers to an alkyl group substituted with anaryl group. Typically, aralkyl groups employed in compounds of thepresent invention have from 1 to 6 carbon atoms incorporated within thealkyl portion of the aralkyl group. Suitable aralkyl groups employed incompounds of the present invention include, for example, benzyl,picolyl, and the like.

Representative heteroaryl groups include, for example, those shownbelow. These heteroaryl groups can be further substituted and may beattached at various positions as will be apparent to those having skillin the organic and medicinal chemistry arts in conjunction with thedisclosure herein.

Representative heteroaryls include, for example, imidazolyl, pyridyl,thiazolyl, triazolyl benzimidazolyl, benzothiazolyl, and benzoxazolyl.

“Biaryl” refers to a group or substituent to which two aryl groups,which are not condensed to each other, are bound. Exemplary biarylcompounds include, for example, phenylbenzene, diphenyldiazene,4-methylthio-1-phenylbenzene, phenoxybenzene, (2-phenylethynyl)benzene,diphenyl ketone, (4-phenylbuta-1,3-diynyl)benzene, phenylbenzylamine,(phenylmethoxy)benzene, and the like. Preferred optionally substitutedbiaryl groups include:2-(phenylamino)-N-[4-(2-phenylethynyl)-phenyl]acetamide,1,4-diphenylbenzene,N-[4-(2-phenylethynyl)phenyl]-2-[benzyl-amino]-acetamide,2-amino-N-[4-(2-phenylethynyl)phenyl]propanamide,2-amino-N-[4-(2-phenyl-ethynyl)phenyl]acetamide,2-(cyclopropylamino)-N-[4-(2-phenylethynyl)-phenyl]-acetamide,2-(ethylamino)-N-[4-(2-phenylethynyl)phenyl]acetamide,2-[(2-methyl-propyl)amino]-N-[4-(2-phenylethynyl)phenyl]acetamide,5-phenyl-2H-benzo-[d]1,3-dioxolene, 2-chloro-1-methoxy-4-phenylbenzene,2-[(imidazolylmethyl)-amino]-N-[4-(2-phenylethynyl)phenyl]acetamide,4-phenyl-1-phenoxybenzene,N-(2-amino-ethyl)-[4-(2-phenylethynyl)phenyl]carboxamide,2-{[(4-fluorophenyl)methyl]-amino}-N-[4-(2-phenylethynyl)phenyl]acetamide,2-{[(4-methylphenyl)methyl]amino}-N-[4-(2-phenyl-ethynyl)phenyl]acetamide,4-phenyl-1-(trifluoromethyl)benzene, 1-butyl-4-phenylbenzene,2-(cyclohexylamino)-N-[4-(2-phenylethynyl)phenyl]acetamide,2-(ethyl-methyl-amino)-N-[4-(2-phenylethynyl)phenyl]acetamide,2-(butylamino)-N-[4-(2-phenyl-ethynyl)-phenyl]acetamide,N-[4-(2-phenylethynyl)phenyl]-2-(4-pyridylamino)-acetamide,N-[4-(2-phenylethynyl)phenyl]-2-(quinuclidin-3-ylamino)acetamide,N-[4-(2-phenyl-ethynyl)phenyl]pyrrolidin-2-ylcarboxamide,2-amino-3-methyl-N-[4-(2-phenyl-ethynyl)-phenyl]butanamide,4-(4-phenylbuta-1,3-diynyl)phenylamine,2-(dimethyl-amino)-N-[4-(4-phenylbuta-1,3-diynyl)phenyl]acetamide,2-(ethylamino)-N-[4-(4-phenylbuta-1,3-diynyl)-phenyl]acetamide,4-ethyl-1-phenylbenzene, 1-[4-(2-phenyl-ethynyl)-phenyl]ethan-1-one,N-(1-carbamoyl-2-hydroxypropyl)[4-(4-phenylbuta-1,3-diynyl)-phenyl]-carbox-amide,N-[4-(2-phenylethynyl)phenyl]propanamide, 4-methoxy-phenyl phenylketone, phenyl-N-benzamide,(tert-butoxy)-N-[(4-phenylphenyl)-methyl]-carboxamide,2-(3-phenyl-phenoxy)ethanehydroxamic acid, 3-phenylphenyl propanoate,1-(4-ethoxyphenyl)-4-methoxybenzene, and[4-(2-phenylethynyl)phenyl]pyrrole.

“Heteroarylaryl” refers to a biaryl group where one of the aryl groupsis a heteroaryl group. Exemplary heteroarylaryl groups include, forexample, 2-phenylpyridine, phenylpyrrole, 3-(2-phenylethynyl)pyridine,phenylpyrazole, 5-(2-phenyl-ethynyl)-1,3-dihydropyrimidine-2,4-dione,4-phenyl-1,2,3-thiadiazole, 2-(2-phenylethynyl)pyrazine,2-phenylthiophene, phenylimidazole, 3-(2-piperazinyl-phenyl)-furan,3-(2,4-dichlorophenyl)-4-methylpyrrole, and the like. Preferredoptionally substituted heteroarylaryl groups include:5-(2-phenylethynyl)pyrimidine-2-ylamine, 1-methoxy-4-(2-thienyl)benzene,1-methoxy-3-(2-thienyl)benzene, 5-methyl-2-phenyl-pyridine,5-methyl-3-phenylisoxazole, 2-[3-(trifluoromethyl)phenyl]furan,3-fluoro-5-(2-furyl)-2-methoxy-1-prop-2-enylbenzene,(hydroxyimino)(5-phenyl(2-thienyl))-methane,5-[(4-methylpiperazinyl)methyl]-2-phenylthiophene,2-(4-ethylphenyl)-thiophene, 4-methyl-thio-1-(2-thienyl)benzene,2-(3-nitrophenyl)thiophene,(tert-butoxy)-N-[(5-phenyl-(3-pyridyl))methyl]carboxamide,hydroxy-N-[(5-phenyl(3-pyridyl))methyl]-amide,2-(phenyl-methylthio)pyridine, and benzylimidazole.

“Heteroarylheteroaryl” refers to a biaryl group where both of the arylgroups is a heteroaryl group. Exemplary heteroarylheteroaryl groupsinclude, for example, 3-pyridylimidazole, 2-imidazolylpyrazine, and thelike. Preferred optionally substituted heteroarylheteroaryl groupsinclude: 2-(4-piperazinyl-3-pyridyl)furan,diethyl-(3-pyrazin-2-yl(4-pyridyl))amine, anddimethyl{2-[2-(5-methylpyrazin-2-yl)ethynyl](4-pyridyl)}amine.

“Optionally substituted” or “substituted” refers to the replacement ofhydrogen with one or more monovalent or divalent radical. Suitablesubstitution groups include, for example, hydroxyl, nitro, amino, imino,cyano, halo, thio, sulfonyl, thioamido, amidino, imidino, oxo,oxamidino, methoxamidino, imidino, guanidino, sulfonamido, carboxyl,formyl, alkyl, substituted alkyl, haloalkyl, alkyamino, haloalkylamino,alkoxy, haloalkoxy, alkoxy-alkyl, alkylcarbonyl, aminocarbonyl,arylcarbonyl, aralkylcarbonyl, heteroarylcarbonyl,heteroaralkyl-carbonyl, alkylthio, aminoalkyl, cyanoalkyl, aryl, benzyl,pyridyl, pyrazolyl, pyrrole, thiophene, imidazolyl, and the like.

The substitution group can itself be substituted. The group substitutedonto the substitution group can be carboxyl, halo, nitro, amino, cyano,hydroxyl, alkyl, alkoxy, aminocarbonyl, —SR, thioamido, —SO₃H, —SO₂R, orcycloalkyl, where R is typically hydrogen, hydroxyl or alkyl.

When the substituted substituent includes a straight chain group, thesubstitution can occur either within the chain (e.g., 2-hydroxypropyl,2-aminobutyl, and the like) or at the chain terminus (e.g.,2-hydroxyethyl, 3-cyanopropyl, and the like). Substituted substituentscan be straight chain, branched or cyclic arrangements of covalentlybonded carbon or heteroatoms.

Representative substituted aminocarbonyl groups include, for example,those shown below. These can be further substituted by heterocyclylgroups and heteroaryl groups as will be apparent to those having skillin the organic and medicinal chemistry arts in conjunction with thedisclosure herein. Preferred aminocarbonyl groups include:N-(2-cyanoethyl)carboxamide, N-(3-methoxypropyl)carboxamide,N-cyclopropylcarboxamide, N-(2-hydroxy-isopropyl)carboxamide, methyl2-carbonylamino-3-hydroxypropanoate, N-(2-hydroxypropyl)carboxamide,N-(2-hydroxy-isopropyl)carboxamide,N-[2-hydroxy-1-(hydroxymethyl)ethyl]carboxamide,N-(2-carbonylaminoethyl)acetamide, N-(2-(2-pyridyl)ethyl)carboxamide,N-(2-pyridylmethyl)carboxamide, N-(oxolan-2-ylmethyl)-carboxamide,N-(4-hydroxypyrrolidin-2-yl)carboxamide,N-[2-(2-hydroxyethoxy)ethyl]-carboxamide,N-(4-hydroxycyclohexyl)carboxamide,N-[2-(2-oxo-4-imidazolinyl)ethyl]-carboxamide,N-(carbonylaminomethyl)acetamide, N-(3-pyrrolidinylpropyl)carboxamide,N-[1-(carbonylaminomethyl)pyrrolidin-3-yl]acetamide,N-(2-morpholin-4-ylethyl)carboxamide,N-[3-(2-oxopyrrolidinyl)propyl]carboxamide,4-methyl-2-oxopiperazinecarbaldehyde,N-(2-hydroxy-3-pyrrolidinylpropyl)carboxamide,N-(2-hydroxy-3-morpholin-4-ylpropyl)carboxamide,N-{2-[(5-cyano-2-pyridyl)amino]ethyl}carboxamide,3-(dimethylamino)pyrrolidinecarbaldehyde,N-[(5-methylpyrazin-2-yl)methyl]carboxamide,2,2,2-trifluoro-N-(1-formylpyrrolidin-3-yl)acetamide,

Representative substituted alkoxycarbonyl groups include, for example,those shown below. These alkoxycarbonyl groups can be furthersubstituted as will be apparent to those having skill in the organic andmedicinal chemistry arts in conjunction with the disclosure herein.

Representative substituted alkoxycarbonyl groups include, for example,those shown below. These alkoxycarbonyl groups can be furthersubstituted as will be apparent to those having skill in the organic andmedicinal chemistry arts in conjunction with the disclosure herein.

The term “protected” with respect to hydroxyl groups, amine groups, andsulfhydryl groups refers to forms of these functionalities which areprotected from undesirable reaction with a protecting group known tothose skilled in the art such as those set forth in Protective Groups inOrganic Synthesis, Greene, T. W.; Wuts, P. G. M., John Wiley & Sons, NewYork, N.Y., (3rd Edition, 1999) which can be added or removed using theprocedures set forth therein. Examples of protected hydroxyl groupsinclude, but are not limited to, silyl ethers such as those obtained byreaction of a hydroxyl group with a reagent such as, but not limited to,t-butyldimethyl-chlorosilane, trimethylchlorosilane,triisopropylchlorosilane, triethylchlorosilane; substituted methyl andethyl ethers such as, but not limited to methoxymethyl ether,methythiomethyl ether, benzyloxymethyl ether, t-butoxymethyl ether,2-methoxyethoxymethyl ether, tetrahydropyranyl ethers, 1-ethoxyethylether, allyl ether, benzyl ether; esters such as, but not limited to,benzoylformate, formate, acetate, trichloroacetate, andtrifluoroacetate. Examples of protected amine groups include, but arenot limited to, amides such as, formamide, acetamide,trifluoroacetamide, and benzamide; imides, such as phthalimide, anddithiosuccinimide; and others. Examples of protected sulfhydryl groupsinclude, but are not limited to, thioethers such as S-benzyl thioether,and S-4-picolyl thioether; substituted S-methyl derivatives such ashemithio, dithio and aminothio acetals; and others.

“Carboxy-protecting group” refers to a carbonyl group which has beenesterified with one of the commonly used carboxylic acid protectingester groups employed to block or protect the carboxylic acid functionwhile reactions involving other functional sites of the compound arecarried out. In addition, a carboxy protecting group can be attached toa solid support whereby the compound remains connected to the solidsupport as the carboxylate until cleaved by hydrolytic methods torelease the corresponding free acid. Representative carboxy-protectinggroups include, for example, alkyl esters, secondary amides and thelike.

Certain compounds of the invention comprise asymmetrically substitutedcarbon atoms. Such asymmetrically substituted carbon atoms can result inthe compounds of the invention comprising mixtures of stereoisomers at aparticular asymmetrically substituted carbon atom or a singlestereoisomer. As a result, racemic mixtures, mixtures of diastereomers,as well as single diastereomers of the compounds of the invention areincluded in the present invention. The terms “S” and “R” configuration,as used herein, are as defined by the IUPAC 1974 “RECOMMENDATIONS FORSECTION E, FUNDAMENTAL STEREOCHEMISTRY, ” Pure Appl. Chem. 45:13-30,1976. The terms α and β are employed for ring positions of cycliccompounds. The α-side of the reference plane is that side on which thepreferred substituent lies at the lower numbered position. Thosesubstituents lying on the opposite side of the reference plane areassigned β descriptor. It should be noted that this usage differs fromthat for cyclic stereoparents, in which “α” means “below the plane” anddenotes absolute configuration. The terms α and β configuration, as usedherein, are as defined by the “Chemical Abstracts Index Guide,” AppendixIV, paragraph 203, 1987.

As used herein, the term “pharmaceutically acceptable salts” refers tothe nontoxic acid or alkaline earth metal salts of the pyrimidinecompounds of the invention. These salts can be prepared in situ duringthe final isolation and purification of the pyrimidine compounds, or byseparately reacting the base or acid functions with a suitable organicor inorganic acid or base, respectively. Representative salts include,but are not limited to, the following: acetate, adipate, alginate,citrate, aspartate, benzoate, benzenesulfonate, bisulfate, butyrate,camphorate, camphorsulfonate, digluconate, cyclopentanepropionate,dodecylsulfate, ethanesulfonate, glucoheptanoate, glycerophosphate,hemi-sulfate, heptanoate, hexanoate, fumarate, hydrochloride,hydrobromide, hydroiodide, 2-hydroxyethanesulfonate, lactate, maleate,methanesulfonate, nicotinate, 2-naphth-alenesulfonate, oxalate, pamoate,pectinate, persulfate, 3-phenylproionate, picrate, pivalate, propionate,succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, andundecanoate. Also, the basic nitrogen-containing groups can bequaternized with such agents as alkyl halides, such as methyl, ethyl,propyl, and butyl chloride, bromides, and iodides; dialkyl sulfates likedimethyl, diethyl, dibutyl, and diamyl sulfates, long chain halides suchas decyl, lauryl, myristyl, and stearyl chlorides, bromides and iodides,aralkyl halides like benzyl and phenethyl bromides, and others. Water oroil-soluble or dispersible products are thereby obtained.

Examples of acids that may be employed to form pharmaceuticallyacceptable acid addition salts include such inorganic acids ashydrochloric acid, hydroboric acid, nitric acid, sulfuric acid andphosphoric acid and such organic acids as formic acid, acetic acid,trifluoroacetic acid, fumaric acid, tartaric acid, oxalic acid, maleicacid, methanesulfonic acid, succinic acid, malic acid, methanesulfonicacid, benzenesulfonic acid, and p-toluenesulfonic acid, citric acid, andacidic amino acids such as aspartic acid and glutamic acid.

Basic addition salts can be prepared in situ during the final isolationand purification of the pyrimidine compounds, or separately by reactingcarboxylic acid moieties with a suitable base such as the hydroxide,carbonate or bicarbonate of a pharmaceutically acceptable metal cationor with ammonia, or an organic primary, secondary or tertiary amine.Pharmaceutically acceptable salts include, but are not limited to,cations based on the alkali and alkaline earth metals, such as sodium,lithium, potassium, calcium, magnesium, aluminum salts and the like, aswell as nontoxic ammonium, quaternary ammonium, and amine cations,including, but not limited to ammonium, tetramethylammonium,tetraethylammonium, methylamine, dimethylamine, trimethylamine,triethylamine, ethylamine, and the like. Other representative organicamines useful for the formation of base addition salts includediethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine,pyridine, picoline, triethanolamine and the like, and basic amino acidssuch as arginine, lysine and ornithine.

As used herein, the term “pharmaceutically acceptable ester” refers toesters which hydrolyze in vivo and include those that break down readilyin the human body to leave the parent compound or a salt thereof.Suitable ester groups include, for example, those derived frompharmaceutically acceptable aliphatic carboxylic acids, particularlyalkanoic, alkenoic, cycloalkanoic and alkanedioic acids, in which eachalkyl or alkenyl moiety advantageously has not more than 6 carbon atoms.Representative examples of particular esters include, but are notlimited to, formates, acetates, propionates, butyrates, acrylates andethylsuccinates.

The term “pharmaceutically acceptable prodrugs” as used herein refers tothose prodrugs of the compounds of the present invention which are,within the scope of sound medical judgment, suitable for use in contactwith the tissues of humans and lower animals without undue toxicity,irritation, allergic response, and the like, commensurate with areasonable benefit/risk ratio, and effective for their intended use, aswell as the zwitterionic forms, where possible, of the compounds of theinvention. The term “prodrug” refers to compounds that are rapidlytransformed in vivo to yield the parent compound of the above formula,for example by hydrolysis in blood. A thorough discussion is provided inHiguchi, T., and V. Stella, “Pro-drugs as Novel Delivery Systems,”A.C.S. Symposium Series 14, and in “Bioreversible Carriers in DrugDesign,” in Edward B. Roche (ed.), American Pharmaceutical Association,Pergamon Press, 1987, both of which are incorporated herein byreference.

“Treating” within the context of the instant invention, means analleviation of symptoms associated with a disorder or disease, or haltof further progression or worsening of those symptoms, or prevention orprophylaxis of the disease or disorder. For example, within the contextof treating patients in need of an inhibitor of PI3K, successfultreatment may include a reduction in the proliferation of capillariesfeeding a tumor or diseased tissue, an alleviation of symptoms relatedto a cancerous growth or tumor, proliferation of capillaries, ordiseased tissue, a halting in capillary proliferation, or a halting inthe progression of a disease such as cancer or in the growth ofcancerous cells. Treatment may also include administering thepharmaceutical formulations of the present invention in combination withother therapies. For example, the compounds and pharmaceuticalformulations of the present invention may be administered before,during, or after surgical procedure and/or radiation therapy. Thecompounds of the invention can also be administered in conjunction withother anti-cancer drugs including those used in antisense and genetherapy.

As used herein, “limit”, “treat” and “treatment” are interchangeableterms as are “limiting” and “treating” and, as used herein, includepreventative (e.g., prophylactic) and palliative treatment or the act ofproviding preventative or palliative treatment.

The term “PI3K-mediated disorder” refers to a disorder that can bebeneficially treated by the inhibition of PI3K.

The term “cellular proliferative diseases” refers to diseases including,for example, cancer, tumor, hyperplasia, restenosis, cardiachypertrophy, immune disorder and inflammation.

The term “cancer” refers to cancer diseases that can be beneficiallytreated by the inhibition of PI3K, including, for example, lung andbronchus; prostate; breast; pancreas; colon and rectum; thyroid; liverand intrahepatic bile duct; hepatocellular; gastric;glioma/glioblastoma; endometrial; melanoma; kidney and renal pelvis;urinary bladder; uterine corpus; uterine cervix; ovary; multiplemyeloma; esophagus; acute myelogenous leukemia; chronic myelogenousleukemia; lymphocytic leukemia; myeloid leukemia; brain; oral cavity andpharynx; larynx; small intestine; non-Hodgkin lymphoma; melanoma; andvillous colon adenoma.

The PI3K inhibitors of this invention, as described herein, can beadministered in the form of acid addition salts. The salts areconveniently formed by reacting a compound, if basic, with a suitableacid, such as have been described above. The salts are quickly formed inhigh yields at moderate temperatures, and often are prepared by merelyisolating the compound from a suitable acidic wash as the final step ofthe synthesis. The salt-forming acid is dissolved in an appropriateorganic solvent, or aqueous organic solvent, such as an alkanol, ketoneor ester. On the other hand, if the compound of this invention isdesired in the free base form, it is isolated from a basic final washstep, according to the usual practice. A preferred technique forpreparing hydrochlorides is to dissolve the free base in a suitablesolvent and dry the solution thoroughly, as over molecular sieves,before bubbling hydrogen chloride gas through it. It will also berecognized that it is possible to administer amorphous forms of the PI3Kinhibitors.

The invention also provides isotopically-labeled PI3K inhibitors, whichare structurally identical to those disclosed above, but for the factthat one or more atoms are replaced by an atom having an atomic mass ormass number different from the atomic mass or mass number usually foundin nature. Examples of isotopes that can be incorporated into compounds,of the invention include isotopes of hydrogen, carbon, nitrogen, oxygen,phosphorous, sulfur, fluorine and chlorine, such as ²H, ³H, ¹³C, ¹⁴C,¹⁵N, ¹⁸O, ¹⁷O, ³¹P, ³²P, ³⁵S, ¹⁸F and ³⁶Cl, respectively. Compounds ofthe present invention, prodrugs thereof, and pharmaceutically acceptablesalts of said compounds and of said prodrugs which contain theaforementioned isotopes and/or other isotopes of other atoms are withinthe scope of this invention. Certain isotopically labeled compounds ofthe present invention, for example those into which radioactive isotopessuch as ³H and ¹⁴C are incorporated, are useful in drug and/or substratetissue distribution assays. Tritiated, i.e., ³H, and carbon-14, i.e.,¹⁴C, isotopes are particularly preferred for their ease of preparationand detectability. Further, substitution with heavier isotopes such asdeuterium, i.e., ²H, may afford certain therapeutic advantages resultingfrom greater metabolic stability, for example increased in vivohalf-life or reduced dosage requirements and, hence, may be preferred insome circumstances. Isotopically labeled compounds of this invention andprodrugs thereof can generally be prepared by carrying out known orreferenced procedures and by substituting a readily availableisotopically labeled reagent for a non-isotopically labeled reagent.

The compounds of the invention are useful in vitro or in vivo ininhibiting the growth of cancer cells. The compounds may be used aloneor in compositions together with a pharmaceutically acceptable carrieror excipient. Pharmaceutical compositions of the present inventioncomprise a therapeutically effective amount of a phosphatidylinositol3-kinase inhibitor compound described herein formulated together withone or more pharmaceutically acceptable carriers. As used herein, theterm “pharmaceutically acceptable carrier” means a non-toxic, inertsolid, semi-solid or liquid filler, diluent, encapsulating material orformulation auxiliary of any type. Some examples of materials which canserve as pharmaceutically acceptable carriers are sugars such aslactose, glucose and sucrose; starches such as corn starch and potatostarch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethyl cellulose and cellulose acetate; powdered tragacanth;malt; gelatin; talc; excipients such as cocoa butter and suppositorywaxes; oils such as peanut oil, cottonseed oil; safflower oil; sesameoil; olive oil; corn oil and soybean oil; glycols; such a propyleneglycol; esters such as ethyl oleate and ethyl laurate; agar; bufferingagents such as magnesium hydroxide and aluminum hydroxide; alginic acid;pyrogen-free water; isotonic saline; Ringer's solution; ethyl alcohol,and phosphate buffer solutions, as well as other non-toxic compatiblelubricants such as sodium lauryl sulfate and magnesium stearate, as wellas coloring agents, releasing agents, coating agents, sweetening,flavoring and perfuming agents, preservatives and antioxidants can alsobe present in the composition, according to the judgment of theformulator. Other suitable pharmaceutically acceptable excipients aredescribed in “Remington's Pharmaceutical Sciences,” Mack Pub. Co., NewJersey, 1991, incorporated herein by reference.

The compounds of the present invention may be administered to humans andother animals orally, parenterally, sublingually, by aerosolization orinhalation spray, rectally, intracistemally, intravaginally,intraperitoneally, bucally, or topically in dosage unit formulationscontaining conventional nontoxic pharmaceutically acceptable carriers,adjuvants, and vehicles as desired. Topical administration may alsoinvolve the use of transdermal administration such as transdermalpatches or ionophoresis devices. The term parenteral as used hereinincludes subcutaneous injections, intravenous, intramuscular,intrasternal injection, or infusion techniques.

Methods of formulation are well known in the art and are disclosed, forexample, in Remington: The Science and Practice of Pharmacy, MackPublishing Company, Easton, Pa., 19th Edition (1995). Pharmaceuticalcompositions for use in the present invention can be in the form ofsterile, non-pyrogenic liquid solutions or suspensions, coated capsules,suppositories, lyophilized powders, transdermal patches or other formsknown in the art.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-propanediol or1,3-butanediol. Among the acceptable vehicles and solvents that may beemployed are water, Ringer's solution, U.S.P. and isotonic sodiumchloride solution. In addition, sterile, fixed oils are conventionallyemployed as a solvent or suspending medium. For this purpose any blandfixed oil may be employed including synthetic mono- or di-glycerides. Inaddition, fatty acids such as oleic acid find use in the preparation ofinjectables. The injectable formulations can be sterilized, for example,by filtration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a drug, it is often desirable to slowthe absorption of the drug from subcutaneous or intramuscular injection.This may be accomplished by the use of a liquid suspension ofcrystalline or amorphous material with poor water solubility. The rateof absorption of the drug then depends upon its rate of dissolutionwhich, in turn, may depend upon crystal size and crystalline form.Alternatively, delayed absorption of a parenterally administered drugform may be accomplished by dissolving or suspending the drug in an oilvehicle. Injectable depot forms are made by forming microencapsulematrices of the drug in biodegradable polymers such aspolylactide-polyglycolide. Depending upon the ratio of drug to polymerand the nature of the particular polymer employed, the rate of drugrelease can be controlled. Examples of other biodegradable polymersinclude poly(orthoesters) and poly(anhydrides). Depot injectableformulations may also be prepared by entrapping the drug in liposomes ormicroemulsions, which are compatible with body tissues.

Compositions for rectal or vaginal administration are preferablysuppositories which can be prepared by mixing the compounds of thisinvention with suitable non-irritating excipients or carriers such ascocoa butter, polyethylene glycol or a suppository wax which are solidat ambient temperature but liquid at body temperature and therefore meltin the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,acetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like.

The solid dosage forms of tablets, dragees, capsules, pills, andgranules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally contain opacifying agents and can also be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions that can be usedinclude polymeric substances and waxes.

The active compounds can also be in micro-encapsulated form with one ormore excipients as noted above. The solid dosage forms of tablets,dragees, capsules, pills, and granules can be prepared with coatings andshells such as enteric coatings, release controlling coatings and othercoatings well known in the pharmaceutical formulating art. In such soliddosage forms the active compound may be admixed with at least one inertdiluent such as sucrose, lactose or starch. Such dosage forms may alsocomprise, as is normal practice, additional substances other than inertdiluents, e.g., tableting lubricants and other tableting aids such amagnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets and pills, the dosage forms may also comprisebuffering agents. They may optionally contain opacifying agents and canalso be of a composition that they release the active ingredient(s)only, or preferentially, in a certain part of the intestinal tract,optionally, in a delayed manner. Examples of embedding compositions thatcan be used include polymeric substances and waxes.

Liquid dosage forms for oral administration include pharmaceuticallyacceptable emulsions, microemulsions, solutions, suspensions, syrups andelixirs. In addition to the active compounds, the liquid dosage formsmay contain inert diluents commonly used in the art such as, forexample, water or other solvents, solubilizing agents and emulsifierssuch as ethyl alcohol, isopropyl alcohol, ethyl carbonate, EtOAc, benzylalcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol,dimethylformamide, oils (in particular, cottonseed, groundnut, corn,germ, olive, castor, and sesame oils), glycerol, tetrahydrofurfurylalcohol, polyethylene glycols and fatty acid esters of sorbitan, andmixtures thereof. Besides inert diluents, the oral compositions can alsoinclude adjuvants such as wetting agents, emulsifying and suspendingagents, sweetening, flavoring, and perfuming agents.

Dosage forms for topical or transdermal administration of a compound ofthis invention include ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants or patches. The active componentis admixed under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulations, ear drops, and the like are also contemplatedas being within the scope of this invention.

The ointments, pastes, creams and gels may contain, in addition to anactive compound of this invention, excipients such as animal andvegetable fats, oils, waxes, paraffins, starch, tragacanth, cellulosederivatives, polyethylene glycols, silicones, bentonites, silicic acid,talc and zinc oxide, or mixtures thereof.

Compositions of the invention may also be formulated for delivery as aliquid aerosol or inhalable dry powder. Liquid aerosol formulations maybe nebulized predominantly into particle sizes that can be delivered tothe terminal and respiratory bronchioles.

Aerosolized formulations of the invention may be delivered using anaerosol forming device, such as a jet, vibrating porous plate orultrasonic nebulizer, preferably selected to allow the formation of anaerosol particles having with a mass medium average diameterpredominantly between 1 to 5 μm. Further, the formulation preferably hasbalanced osmolarity ionic strength and chloride concentration, and thesmallest aerosolizable volume able to deliver effective dose of thecompounds of the invention to the site of the infection. Additionally,the aerosolized formulation preferably does not impair negatively thefunctionality of the airways and does not cause undesirable sideeffects.

Aerosolization devices suitable for administration of aerosolformulations of the invention include, for example, jet, vibratingporous plate, ultrasonic nebulizers and energized dry powder inhalers,that are able to nebulize the formulation of the invention into aerosolparticle size predominantly in the size range from 1-5 μm. Predominantlyin this application means that at least 70% but preferably more than 90%of all generated aerosol particles are within 1-5 μm range. A jetnebulizer works by air pressure to break a liquid solution into aerosoldroplets. Vibrating porous plate nebulizers work by using a sonic vacuumproduced by a rapidly vibrating porous plate to extrude a solventdroplet through a porous plate. An ultrasonic nebulizer works by apiezoelectric crystal that shears a liquid into small aerosol droplets.A variety of suitable devices are available, including, for example,AERONEB and AERODOSE vibrating porous plate nebulizers (AeroGen, Inc.,Sunnyvale, Calif.), SIDESTREAM nebulizers (Medic-Aid Ltd., West Sussex,England), PARI LC and PART LC STAR jet nebulizers (Pari RespiratoryEquipment, Inc., Richmond, Va.), and AEROSONIC (DeVilbiss MedizinischeProdukte (Deutschland) GmbH, Heiden, Germany) and ULTRAAIRE (OmronHealthcare, Inc., Vernon Hills, Ill.) ultrasonic nebulizers.

Compounds of the invention may also be formulated for use as topicalpowders and sprays that can contain, in addition to the compounds ofthis invention, excipients such as lactose, talc, silicic acid, aluminumhydroxide, calcium silicates and polyamide powder, or mixtures of thesesubstances. Sprays can additionally contain customary propellants suchas chlorofluorohydrocarbons.

Transdermal patches have the added advantage of providing controlleddelivery of a compound to the body. Such dosage forms can be made bydissolving or dispensing the compound in the proper medium. Absorptionenhancers can also be used to increase the flux of the compound acrossthe skin. The rate can be controlled by either providing a ratecontrolling membrane or by dispersing the compound in a polymer matrixor gel. The compounds of the present invention can also be administeredin the form of liposomes. As is known in the art, liposomes aregenerally derived from phospholipids or other lipid substances.Liposomes are formed by mono- or multi-lamellar hydrated liquid crystalsthat are dispersed in an aqueous medium. Any non-toxic, physiologicallyacceptable and metabolizable lipid capable of forming liposomes can beused. The present compositions in liposome form can contain, in additionto a compound of the present invention, stabilizers, preservatives,excipients, and the like. The preferred lipids are the phospholipids andphosphatidyl cholines (lecithins), both natural and synthetic. Methodsto form liposomes are known in the art. See, for example, Prescott(ed.), “Methods in Cell Biology,” Volume XIV, Academic Press, New York,1976, p. 33 et seq.

Effective amounts of the compounds of the invention generally includeany amount sufficient to detectably inhibit PI3K activity by any of theassays described herein, by, other PI3K activity assays known to thosehaving ordinary skill in the art, or by detecting an inhibition oralleviation of symptoms of cancer. The amount of active ingredient thatmay be combined with the carrier materials to produce a single dosageform will vary depending upon the host treated and the particular modeof administration. It will be understood, however, that the specificdose level for any particular patient will depend upon a variety offactors including the activity of the specific compound employed, theage, body weight, general health, sex, diet, time of administration,route of administration, rate of excretion, drug combination, and theseverity of the particular disease undergoing therapy. Thetherapeutically effective amount for a given situation can be readilydetermined by routine experimentation and is within the skill andjudgment of the ordinary clinician.

According to the methods of treatment of the present invention, tumorgrowth is reduced or prevented in a patient such as a human or lowermammal by administering to the patient a therapeutically effectiveamount of a compound of the invention, in such amounts and for such timeas is necessary to achieve the desired result. By a “therapeuticallyeffective amount” of a compound of the invention is meant a sufficientamount of the compound to treat tumor growth, at a reasonablebenefit/risk ratio applicable to any medical treatment. It will beunderstood, however, that the total daily usage of the compounds andcompositions of the present invention will be decided by the attendingphysician within the scope of sound medical judgment. The specifictherapeutically effective dose level for any particular patient willdepend upon a variety of factors including the disorder being treatedand the severity of the disorder; the activity of the specific compoundemployed; the specific composition employed; the age, body weight,general health, sex and diet of the patient, the time of administration,route of administration, and rate of excretion of the specific compoundemployed; the duration of the treatment; drugs used in combination orcoincidental with the specific compound employed; and like factors wellknown in the medical arts.

For purposes of the present invention, a therapeutically effective dosewill generally be a total daily dose administered to a host in single ordivided doses may be in amounts, for example, of from 0.001 to 1000mg/kg body weight daily and more preferred from 1.0 to 30 mg/kg bodyweight daily. Dosage unit compositions may contain such amounts ofsubmultiples thereof to make up the daily, dose. In general, treatmentregimens according to the present invention comprise administration to apatient in need of such treatment from about 10 mg to about 2000 mg ofthe compound(s) of this invention per day in single or multiple doses.

In another aspect of the invention, kits that include one or morecompounds of the invention are provided. Representative kits include aPI3K inhibitor compound of the invention (e.g., a compound of formulas(I)-(III)) and a package insert or other labeling including directionsfor treating a cellular proliferative disease by administering an PI3Kinhibitory amount of the compound.

The term “kit” as used herein comprises a container for containing thepharmaceutical compositions and may also include divided containers suchas a divided bottle or a divided foil packet. The container can be inany conventional shape or form as known in the art which is made of apharmaceutically acceptable material, for example a paper or cardboardbox, a glass or plastic bottle or jar, a resealable bag (for example, tohold a “refill” of tablets for placement into a different container), ora blister pack with individual doses for pressing out of the packaccording to a therapeutic schedule. The container employed can dependon the exact dosage form involved, for example a conventional cardboardbox would not generally be used to hold a liquid suspension. It isfeasible that more than one container can be used together in a singlepackage to market a single dosage form. For example, tablets may becontained in a bottle which is in turn contained within a box.

An example of such a kit is a so-called blister pack. Blister packs arewell known in the packaging industry and are being widely used for thepackaging of pharmaceutical unit dosage forms (tablets, capsules, andthe like). Blister packs generally consist of a sheet of relativelystiff material covered with a foil of a preferably transparent plasticmaterial. During the packaging process, recesses are formed in theplastic foil. The recesses have the size and shape of individual tabletsor capsules to be packed or may have the size and shape to accommodatemultiple tablets and/or capsules to be packed. Next, the tablets orcapsules are placed in the recesses accordingly and the sheet ofrelatively stiff material is sealed against the plastic foil at the faceof the foil which is opposite from the direction in which the recesseswere formed. As a result, the tablets or capsules are individuallysealed or collectively sealed, as desired, in the recesses between theplastic foil and the sheet. Preferably the strength of the sheet is suchthat the tablets or capsules can be removed from the blister pack bymanually applying pressure on the recesses whereby an opening is formedin the sheet at the place of the recess. The tablet or capsule can thenbe removed via said opening.

It maybe desirable to provide a written memory aid, where the writtenmemory aid is of the type containing information and/or instructions forthe physician, pharmacist or other health care provider, or subject,e.g., in the form of numbers next to the tablets or capsules whereby thenumbers correspond with the days of the regimen which the tablets orcapsules so specified should be ingested or a card which contains thesame type of information. Another example of such a memory aid is acalendar printed on the card, e.g., as follows “First Week, Monday,Tuesday,” “Second Week, Monday, Tuesday, . . . .” Other variations ofmemory aids will be readily apparent. A “daily dose” can be a singletablet or capsule or several tablets or capsules to be taken on a givenday. When the kit contains separate compositions, a daily dose of one ormore compositions of the kit can consist of one tablet or capsule whilea daily dose of another one or more compositions of the kit can consistof several tablets or capsules.

Another specific embodiment of a kit is a dispenser designed to dispensethe daily doses one at a time in the order of their intended use.Preferably, the dispenser is equipped with a memory-aid, so as tofurther facilitate compliance with the regimen. An example of such amemory-aid is a mechanical counter, which indicates the number of dailydoses that has been dispensed. Another example of such a memory-aid is abattery-powered micro-chip memory coupled with a liquid crystal readout,or audible reminder signal which, for example, reads out the date thatthe last daily dose has been taken and/or reminds one when the next doseis to be taken.

The kits of the present invention may also comprise, in addition to aPI3K inhibitor, one or more additional pharmaceutically activecompounds. Preferably, the additional compound is another PI3K inhibitoror another compound useful to treat cancer, angiogenesis, or tumorgrowth. The additional compounds may be administered in the same dosageform as the PI3K inhibitor or in different dosage forms. Likewise, theadditional compounds can be administered at the same time as the PI3Kinhibitor or at different times.

The present invention will be understood more readily by reference tothe following examples, which are provided by way of illustration andare not intended to be limiting of the present invention.

EXAMPLES

Referring to the examples that follow, compounds of the presentinvention were synthesized using the methods described herein, or othermethods, which are known in the art.

The compounds and/or intermediates were characterized by highperformance liquid chromatography (HPLC) using a Waters Milleniumchromatography system with a 2695 Separation Module (Milford, Mass.).The analytical columns were Alltima C-18 reversed phase, 4.6×50 mm, flow2.5 mL/min, from Alltech (Deerfield, Ill.). A gradient elution was used,typically starting with 5% acetonitrile/95% water and progressing to100% acetonitrile over a period of 40 minutes. All solvents contained0.1% trifluoroacetic acid (TFA). Compounds were detected by ultravioletlight (UV) absorption at either 220 or 254 rim. HPLC solvents were fromBurdick and Jackson (Muskegan, Mich.), or Fisher Scientific (Pittsburgh,Pa.). In some instances, purity was assessed by thin layerchromatography (TLC) using glass or plastic backed silica gel plates,such as, for example, Baker-Flex Silica Gel 1B2-F flexible sheets. TLCresults were readily detected visually under ultraviolet light, or byemploying well known iodine vapor and other various staining techniques.

Mass spectrometric analysis was performed on one of two LCMSinstruments: a Waters System (Alliance HT HPLC and a Micromass ZQ massspectrometer; Column: Eclipse XDB-C18, 2.1×50 mm; solvent system: 5-95%(or 35-95%, or 65-95% or 95-95%) acetonitrile in water with 0.05% TFA;flow rate 0.8 mL/min; molecular weight range 200-1500; cone Voltage 20V; column temperature 40° C.) or a Hewlett Packard System (Series 1100HPLC; Column: Eclipse XDB-C18, 2.1×50 mm; solvent system: 1-95%acetonitrile in water with 0.05% TFA; flow rate 0.8 mL/min; molecularweight range 150-850; cone Voltage 50 V; column temperature 30° C.). Allmasses were reported as those of the protonated parent ions.

GCMS analysis is performed on a Hewlett Packard instrument (HP6890Series gas chromatograph with a Mass Selective Detector 5973; injectorvolume: 1 μL; initial column temperature: 50° C.; final columntemperature: 250° C.; ramp time: 20 minutes; gas flow rate: 1 mL/min;column: 5% phenyl methyl siloxane, Model No. HP 190915-443, dimensions:30.0 m×25 m×0.25 m).

Nuclear magnetic resonance (NMR) analysis was performed on some of thecompounds with a Varian 300 MHz NMR (Palo Alto, Calif.). The spectralreference was either TMS or the known chemical shift of the solvent.Some compound samples were run at elevated temperatures (e.g., 75° C.)to promote increased sample solubility.

The purity of some of the invention compounds is assessed by elementalanalysis (Desert Analytics, Tucson, Ariz.).

Melting points are determined on a Laboratory Devices Mel-Temp apparatus(Holliston, Mass.).

Preparative separations were carried out using a Flash 40 chromatographysystem and KP-Sil, 60A (Biotage, Charlottesville, Va.), or by flashcolumn chromatography using silica gel (230-400 mesh) packing material,or by HPLC using a Waters 2767 Sample Manager, C-18 reversed phasecolumn, 30×50 mm, flow 75 mL/min. Typical solvents employed for theFlash 40 Biotage system and flash column chromatography weredichloromethane, methanol, ethyl acetate, hexane, acetone, aqueousammonia (or ammonium hydroxide), and triethyl amine. Typical solventsemployed for the reverse phase HPLC were varying concentrations ofacetonitrile and water with 0.1% trifluoroacetic acid.

It should be understood that the organic compounds according to theinvention may exhibit the phenomenon of tautomerism. As the chemicalstructures within this specification can only represent one of thepossible tautomeric forms, it should be understood that the inventionencompasses any tautomeric form of the drawn structure.

It is understood that the invention is not limited to the embodimentsset forth herein for illustration, but embraces all such forms thereofas come within the scope of the above disclosure.

ABBREVIATIONS

ACN Acetonitrile BINAP 2,2′-bis(diphenylphosphino)-1,1′-binapthyl DIEAdiisopropylethylamine DME 1,2-dimethoxyethane DMF N,N-dimethylformamideDPPF 1,1′-bis(diphenylphosphino)ferrocene EtOAc ethyl acetate EtOHethanol MCPBA meta-chloroperoxybenzoic acid NBS N-bromosuccinimide NMPN-methyl-2-pyrrolidone RT room temperature THF tetrahydrofuran

General Methods for Synthesizing PI3K Inhibitor Compounds

Methods for preparing compounds of Formula I and/or II are provided. Themethods include: reacting a 4-halo-2-morpholinopyrimidine with asubstituted pyridinyl or pyrimidinyl group containing a reactive boronicester substituent, in the presence of a palladium catalyst. In oneembodiment, the substituted pyridinyl or pyrimidinyl group containing areactive boronic ester substituent has an —NH₂ group positioned para tothe boronic ester. In another embodiment, the substituted pyridinyl orpyrimidinyl group containing a reactive boronic ester substituent has an—NH₂ group positioned para to the boronic ester and another non-hydrogensubstituent positioned ortho to the boronic ester. In certainembodiments, the non-hydrogen substituent is —CF₃, —CN, —NH₂, halo, orsubstituted or unsubstituted C₁₋₃ alkyl.

In another embodiment, the 4-halo-2-morpholinopyrimidine group is a4-halo-6-heterocyclyl-2-morpholinopyrimidine group. In anotherembodiment, the 4-halo-2-morpholinopyrimidine group is a4-halo-6-heterocyclyloxy-2-morpholinopyrimidine group. In anotherembodiment, the 4-halo-2-morpholinopyrimidine group is a4-halo-6-heteroarylamino-2-morpholinopyrimidine group. In anotherembodiment, the 4-halo-2-morpholinopyrimidine group is4-chloro-2,6-dimorpholinopyrimidine.

In another embodiment, the pyridinyl boronic ester is4-(trifluoromethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine.In another embodiment, the palladium catalyst is Pd(dppf)Cl₂dichloromethane adduct.

In another embodiment, the 4-halo-6-heterocyclyl-2-morpholinopyrimidinegroup is prepared by reacting a heterocyclyl group with a4,6-dihalo-2-morpholinopyrimidine group. In another embodiment, the4-chloro-2,6-dimorpholinopyrimidine group is prepared by reacting4,6-dichloro-2-morpholinopyrimidine with morpholine. In anotherembodiment, the 4,6-dichloro-2-morpholinopyrimidine group is prepared byreacting 2-morpholinopyrimidine-4,6-diol with POCl₃. In anotherembodiment, the 2-morpholinopyrimidine-4,6-diol is prepared by reactingmorpholine-4-carboxamidine with diethyl malonate in the presence of abase, such as sodium ethoxide.

In another embodiment, the substituted pyridinyl or pyrimidinyl groupcontaining a reactive boronic ester substituent is prepared by reactinga substituted pyridinyl or pyrimidinyl group containing a bromosubstituent with a diboronic ester, such as4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane.In another embodiment, the substituted pyridinyl or pyrimidinyl groupcontaining a bromo substituent is prepared by reacting the substitutedpyridinyl or pyrimidinyl group with N-bromosuccinimide (NBS).

Another embodiment of the present invention provides a method ofpreparing a 4-chloro-2,6-dimorpholinopyrimidine comprising reactingmorpholine with 2,4,6-trichloropyrimidine in a suitable solvent. In amore particular embodiment, the solvent is a polar aprotic solvent. Moreparticular still the solvent is THF. In another more particularembodiment, the 4-chloro-2,6-dimorpholinopyrimidine is added over aperiod of at least 10 minutes, or at least 20 minutes, or 30 minutes toa solution comprising morpholine. Alternatively, the morpholine is addedto a solution comprising 4-chloro-2,6-dimorpholinopyrimidine. Moreparticular still, the solution is cooled below 20° C., or below 10° C.,or below 5° C., or below 0° C. More particularly, during or afteraddition of the 4-chloro-2,6-dimorpholinopyrimidine, the solution isallowed to warm to greater than 20° C., or greater than 25° C., orgreater than 30° C. In another embodiment, after the morpholine and4-chloro-2,6-dimorpholinopyrimidine are combined, the solution isquenched by addition of an aqueous solution. More particularly, at least10 hours, or at least 20 hours, or at least 30 hours, or at least 40hours, or at least 50 hours, or about 64 hours after the morpholine and4-chloro-2,6-dimorpholinopyrimidine are combined, the solution isquenched by addition of an aqueous solution. More particularly, afterquenching, the solution is purified by column chromatography. Moreparticular still, the column is silica gel. In another embodiment, the4-chloro-2,6-dimorpholinopyrimidine is reacted with a 2-aminopyridyl or2-aminopyrimidyl moiety to form a compound of Formula III.

Compounds of the invention containing a pyrimidine core, such as thoseof Formula I, may be prepared using a number of methods familiar to oneof skill in the art. In one method, suitably functionalized amines maybe coupled with 4,6-dichloro-2-morpholinopyrimidine by nucleophilicaromatic substitution reactions or by a Buchwald-Hartwig cross-couplingreaction (Hartwig et al., Tetrahedron Letters 36, (1995) 3609), whereinAr represents aryl or heteroaryl moieties. Subsequently, Suzuki coupling(Suzuki et al., Chem. Commun. (1979) 866) to form the final product maybe effected under known conditions such as by treatment withfunctionalized boronic esters as in the following schemes:

From 2,4,6-tribromopyrimidine: SNAr (or Buchwald) reaction offunctionalized arylamines with 2,4,6-tribromopyrimidine gavepreferentially 4-substituted products. Morpholine substitution at2-position followed with Suzuki reaction affords the final pyrimidineanalogs:

Alternatively, multiple Suzuki couplings can be used to afford aryl orheteroaryl groups appended directly to the pyrimidine core at the 4 and6 positions; or an initial Suzuki coupling can be performed followed bya nucleophilic aromatic substitution reaction or a Buchwald-Hartwigcross-coupling reaction, as shown in the following scheme:

More particular syntheses of compounds of the present invention,particularly those of Formula I, II, and III, are provided in thefollowing Methods and Examples:

Method 1 Synthesis of5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidine-2-ylamine

To a dry 500-mL flask was added 2-amino-5-bromopyrimidine (10 g, 57.5mmol), potassium acetate (16.9 g, 172 mmol),4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2yl)-1,3,2-dioxaborolane(16.1 g, 63.0 mmol) and dioxane (300 mL). Argon was bubbled through thesolution for 15 minutes, at which timedichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium (II)dichloromethane adduct (Pd(dppf)Cl₂ CH₂Cl₂) (2.34 g, 2.87 mmol) wasadded. The reaction mixture was refluxed in a 115° C. oil bath for 4hours under argon. After cooling to room temperature, EtOAc (500 mL) wasadded and the resulting slurry was sonicated and filtered. AdditionalEtOAc (500 mL) was used to wash the solid. The combined organic extractswere washed with H₂O (2×300 mL), NaCl_((sat.)) (300 mL), dried overNa₂SO₄, and filtered through a 5 cm pad of silica gel. Additional EtOAcwas used to flush product. After the solvent was concentrated, the crudewas treated with a mixture of 1:3 dichloromethane and hexane (40 mL),filtered and washed with hexane yielding a light yellow solid (8.5 g,75%). LCMS (m/z): 140 (MH⁺ of boronic acid, deriving from producthydrolysis on LC). ¹H NMR (CDCl₃): δ 8.58 (s, 2H), 5.74 (s, 2H), 1.32(s, 12H).

Method 2 Synthesis of 2-Aminomethyl-5-bromopyrimidine

Methylamine (2.0 M in methanol, 40 mL, 80 mmol) was added to5-bromo-2-chloropyrimidine (5.6 g, 29.0 mmol) in a sealable reactionvessel. After allowing to vent for a few minutes, the vessel was sealed,placed behind a safety shield and heated in a 115° C. oil bath for 48hours. Upon cooling the volatiles were removed in vacuo. The materialwas dissolved in CH₂Cl₂ (200 mL) and washed with 1M NaOH (40 mL). Theaqueous layer was extracted further with CH₂Cl₂ (2×50 mL). The combinedorganics were dried over MgSO₄, filtered and concentrated yielding anoff white solid (5.1 g, 93%). LCMS (m/z): 188.0/190.0 (MH⁺).

Synthesis ofmethyl[5-(4,4,5,5-tetramethyl(1,3,2-dioxaborolan-2-yl))pyrimidin-2-yl]amine

To a dry 500 mL flask was added 2-methylamino-5-bromopyrimidine (9.5 g,50.5 mmol), potassium acetate (15.1 g, 154.4 mmol),4,4,5,5,-tetramethyl-2-(4,4,5,5,-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane(14.1 g, 55.5 mmol) and dioxane (280 mL). Argon was bubbled through thesolution for 15 minutes, at which time1,1′-bis(diphenylphosphino)ferrocene palladium(II) chloridedichloromethane adduct (2.05 g, 2.51 mmol) was added. The reaction wasrefluxed in a 115° C. oil bath for 4 hours under argon. After cooling toroom temperature, EtOAc (500 mL) was added and the resulting slurry wassonicated and filtered. Additional EtOAc (500 mL) was used to wash thesolid. The combined organics were washed with H₂O (2×300 mL),NaCl_((sat.)), (300 mL), dried over Na₂SO₄, filtered and the solventswere removed in vacuo. Purification by SiO₂ chromatography (50%EtOAc/hexanes) yielded an off white solid (7.66 g, 64%). LCMS (m/z): 154(MH⁺ of boronic acid, deriving from in situ product hydrolysis on LC).¹H NMR (CDCl₃): δ 8.58 (s, 2H), 5.56 (s, 1H), 3.02 (d, 3H), 1.32 (s,12H).

Method 3 Synthesis of 5-bromo-4-methylpyrimidine-2-ylamine

To a solution of 4-methylpyrimidine-2-ylamine (10.9 g, 100 mmol) inchloroform (400 mL) was added N-bromosuccinimide (17.8 g, 100 mmol). Thesolution was stirred in the dark for 15 hours, at which time it wasadded to CH₂Cl₂ (1400 mL), washed with 1N NaOH (3×200 mL) andNaCl_((sat.)) (100 mL), dried over Na₂SO₄, filtered and concentrated,yielding 5-bromo-4-methylpyrimidine-2-ylamine (18.8 g, 99%). LCMS (m/z):188.0/190.0 (MH⁺). ¹H NMR (CDCl₃): δ 8.22 (s, 1H), 5.02 (bs, 2H), 2.44(s, 3H).

Synthesis of4-methyl-5-(4,4,5,5-tetramethyl(1,3,2-dioxaborolan-2-yl))pyrimidine-2-ylamine

To a dry 1 L flask was added 5-bromo-4-methylpyrimidine-2-ylamine (18.8g, 100 mmol), potassium acetate (29.45 g, 300 mmol),4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane(26.7 g, 105 mmol) and dioxane (500 mL). Argon was bubbled through thesolution for 15 minutes, at which time1,1′-bis(diphenylphosphino)ferrocene palladium(II) chloridedichloromethane adduct (4.07 g, 5 mmol) was added. The reaction wasrefluxed in a 115° C. oil bath for 18 hours under argon. After coolingto room temperature, EtOAc (500 mL) was added and the resulting slurrywas sonicated and filtered. Additional EtOAc (500 mL) was used to washthe solid. The combined organic extracts were washed with H₂O (2×300mL), NaCl_((sat.)) (300 mL), dried over Na₂SO₄, concentrated andpurified by SiO₂ chromatography (EtOAc eluent) yielding 18.1 g of anoff-white solid. By ¹H NMR the material was a 5:1 mixture of boronateester and 4-methylpyrimidine-2-ylamine as a byproduct. The material wasused as is in subsequent Suzuki reactions. LCMS (m/z): 154 (MH⁺ ofboronic acid, deriving from in situ product hydrolysis on LC). ¹H NMR(CDCl₃): δ 8.52 (s, 1H), 5.14 (bs, 2H), 2.56 (d, 3H), 1.32 (s, 12H).

Method 4 Synthesis of 5-bromo-4-(trifluoromethyl)-2-pyridylamine

To a solution of 2-amino-4-trifluoromethylpyridine (10.0 g, 62.1 mmol)in chloroform (200 mL) was added N-bromosuccinimide (12.0 g, 67.4 mmol).The solution was stirred in the dark for 2 hours, at which time it wasadded to CH₂Cl₂ (200 mL) and 1N NaOH (200 mL). Upon mixing, the layerswere separated and the organic layer was washed with NaCl_((sat.)) (100mL), dried over Na₂SO₄, filtered and concentrated. The crude materialwas purified by SiO₂ chromatography (0-5% EtOAc/CH₂Cl₂) yielding 12.0 g(80%) of 5-bromo-4-(trifluoromethyl)-2-pyridylamine LCMS (m/z): 241/243(MH⁺). ¹H NMR (CDCl₃): δ 8.28 (s, 1H), 6.77 (s, 1H), 4.78 (bs, 2H).

Synthesis of5-(4,4,5,5-tetramethyl(1,3,2-dioxaborolan-2-yl))-4-(trifluoromethyl)-2-pyridylamine

To a dry 500 mL flask was added5-bromo-4-(trifluoromethyl)-2-pyridylamine (11.8 g, 49.0 mmol),potassium acetate (14.4 g, 146.9 mmol),4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane(13.6 g, 53.9 mmol) and dioxane (300 mL). Argon was bubbled through thesolution for 15 minutes, at which time1,1′-bis(diphenylphosphino)ferrocene palladium(II) chloridedichloromethane adduct (2.0 g, 2.45 mmol) was added. The reaction wasrefluxed in a 115° C. oil bath for 8 hours under argon. After cooling toroom temperature, the dioxane was removed in vacuo. EtOAc (500 mL) wasadded, and the resulting slurry was sonicated and filtered. AdditionalEtOAc (500 mL) was used to wash the solid. The combined organic extractswere concentrated and the crude material was partially purified by SiO₂chromatography (30-40% EtOAc/Hexanes). Upon removal of solvent, hexanes(75 mL) was added; after sonication, the resulting solid was filteredand dried on a high vacuum for 3 days yielding 2.4 g of an off-whitesolid. By ¹H NMR the material was a 5:1 mixture of boronate ester and2-amino-4-trifluoromethyl pyridine byproduct. The material was used asis in subsequent Suzuki reactions. LCMS (m/z): 207 (MH⁺ of boronic acid,deriving from in situ product hydrolysis on LC). ¹H NMR (CDCl₃): δ 8.50(s, 1H), 6.72 (s, 1H), 4.80 (bs, 2H), 1.34 (s, 12H).

Method 5 Synthesis of 5-bromo-4-(trifluoromethyl)pyrimidin-2-amine

To a solution of 2-amino-4-trifluoromethylpyrimidine (8.0 g, 49.1 mmol)in chloroform (300 mL) was added N-bromosuccinimide (8.9 g, 50 mmol).The solution was stirred in the dark for 16 hours, at which timeadditional N-bromosuccinimide (4.0 g, 22.5 mmol) was added. Afterstirring for an additional 4 hours the solution was added to CH₂Cl₂ (200mL) and 1N NaOH (200 mL). Upon mixing, the layers were separated and theorganic layer was washed with NaCl_((sat.)) (100 mL), dried over Na₂SO₄,filtered and concentrated, yielding 10.9 g (82%) of5-bromo-4-(trifluoromethyl)-2-pyrimidylamine. LCMS (m/z): 242/244 (MH⁺).¹H NMR (CDCl₃): δ 8.52 (s, 1H), 5.38 (bs, 2H).

Synthesis of5-(4,4,5,5-tetramethyl(1,3,2-dioxaborolan-2-yl))-4-(trifluoromethyl)pyrimidine-2-ylamine

To a dry 500 mL flask was added5-bromo-4-(trifluoromethyl)-2-pyrimidylamine (10.1 g, 41.7 mmol),potassium acetate (12.3 g, 125.2 mmol),4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane(11.6 g, 45.9 mmol) and dioxane (150 mL). Argon was bubbled through thesolution for 15 minutes, at which time1,1′-bis(diphenylphosphino)ferrocene palladium (II) chloride (1.7 g, 2.1mmol) was added. The reaction was refluxed in a 115° C. oil bath for 6hours under argon. After cooling to room temperature, the dioxane wasremoved in vacuo. EtOAc (500 mL) was added and the resulting slurry wassonicated and filtered. Additional EtOAc (500 mL) was used to wash thesolid. The combined organic extracts were concentrated and the crudematerial was purified by SiO₂ chromatography (30-40% EtOAc/hexanes)yielding 4.40 g of an off white solid. By ¹H NMR the material was a 1:1mixture of boronate ester and 2-amino-4-trifluoromethylpyrimidinebyproduct. The material was used as is in subsequent Suzuki reactions.LCMS (m/z): 208 (MH⁺ of boronic acid, deriving from in situ producthydrolysis on LC). ¹H NMR (CDCl₃): δ 8.72 (s, 1H), 5.50 (bs, 2H), 1.34(s, 12H).

Method 6 Synthesis of 5-bromo-4-chloro-2-pyridylamine

To a solution of 4-chloro-2-pyridylamine (6.0 g, 46.7 mmol) inchloroform (180 mL) was added N-bromosuccinimide (8.3 g, 46.7 mmol). Thesolution was stirred in the dark for 2 hours, at which time it was addedto CH₂Cl₂ (800 mL) and 1N NaOH (100 mL). Upon mixing, the layers wereseparated and the organic layer was washed with NaCl_((sat.)) (100 mL),dried over Na₂SO₄, filtered and concentrated. The crude material waspurified by SiO₂ chromatography (25-35% EtOAc/hexanes) yielding 3.63 g(38%) of 5-bromo-4-chloro-2-pyridylamine. LCMS (m/z): 206.9/208.9 (MH⁺).¹H NMR (CDCl₃): δ 8.18 (s, 1H), 6.62 (s, 1H), 4.52 (bs, 2H).

Synthesis of4-chloro-5-(4,4,5,5-tetramethyl(1,3,2-dioxaborolan-2-yl))-2-pyridylamine

To a dry 500-mL flask was added 5-bromo-4-chloro 2-pyridylamine (7.3 g,35.8 mmol), potassium acetate (10.3 g, 105 mmol),4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane(10.1 g, 39.8 mmol) and dioxane (150 mL). Argon was bubbled through thesolution for 15 minutes, at which time1,1′-bis(diphenylphosphino)ferrocene palladium(II) chloridedichloromethane adduct (0.85 g, 1.04 mmol) was added. The reaction wasrefluxed in a 115° C. oil bath for 6 hours under argon. After cooling toroom temperature, the dioxane was removed in vacuo. EtOAc (500 mL) wasthen added and the resulting slurry was sonicated and filtered.Additional EtOAc (500 mL) was used to wash the solid. The combinedorganic extracts were concentrated and the crude material was purifiedby SiO₂ chromatography (EtOAc as eluent). Upon removal of solvent, 3:1hexanes/CH₂Cl₂ was added (100 mL). After sonication, the resulting solidwas filtered and concentrated in vacuo yielding 2.8 g of a white solid.By ¹H NMR the material was a 10:1 mixture of boronate ester and2-amino-4-chloropyridine byproduct. The material was used as is insubsequent Suzuki reactions. LCMS (m/z): 173 (MH⁺ of boronic acid,deriving from in situ product hydrolysis on LC). ¹H NMR (CDCl₃): δ 8.36(s, 1H), 6.46 (s, 1H), 4.70 (bs, 2H), 1.38 (s, 12H).

Method 7 Synthesis of 5-bromopyrimidine-2,4-diamine

To a solution of 2,4-diaminopyrimidine (1.0 g, 9.1 mmol) in chloroform(30 mL) was added N-bromosuccinimide (1.62 g, 9.08 mmol). The solutionwas stirred in the dark for 12 hours, at which time it was added toCH₂Cl₂ (150 mL) and 1N NaOH (50 mL). The solid that formed was filtered,rinsed with water and concentrated in vacuo, yielding 1.4 g (74%) of5-bromopyrimidine-2,4-diamine. LCMS (m/z): 189/191 (MH⁺). ¹H NMR(DMSO-_(d6)): δ 7.78 (s, 1H), 6.58 (bs, 2H), 6.08 (bs, 2H).

Synthesis of5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidine-2,4-diamine

To a dry 1 L flask was added 5-bromopyrimidine-2,4-diamine (30.0 g,158.7 mmol), potassium acetate (45.8 g, 466.7 mmol),4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane(51.2 g, 202.2 mmol) and dioxane (500 mL). Argon was bubbled through thesolution for 15 minutes, at which time1,1′-bis(diphenylphosphino)ferrocene palladium(II) chloride (2.5 g, 3.11mmol) was added. The reaction was refluxed in a 115° C. oil bath for 16hours under argon. After cooling to room temperature, the solidinorganic material was filtered, rinsed with EtOAc (1 L). The organicfiltrate was concentrated in vacuo and to the resulting solid was addeddichloromethane (1 L). After sonication the solid was filtered. Thesolid was the debrominated 2,4-diaminopyrimidine. The filtratecontaining desired boronate ester was concentrated in vacuo. To thisresidue was added diethyl ether (100 mL). After sonication, the solutionwas filtered, rinsed with additional diethyl ether (50 mL) and the solidobtained was dried under high vacuum to yield the desired2,4-diaminopyrimidyl-5-boronate ester (10.13 g, 27%). By ¹H NMR thematerial was a 4:1 mixture of 2,4-diaminopyrimidyl-5-boronate ester and2,4-diaminopyrimidine byproduct. The material was used as is insubsequent Suzuki reactions. LCMS (m/z): 155 (MH⁺ of boronic acid,deriving from in situ product hydrolysis on LC). ¹H NMR (CDCl₃+CD₃OD): δ8.16 (s, 1H), 1.34 (s, 12H).

Method 8 Synthesis of 4-methoxypyrimidine-2-ylamine

To a solution of 4,6-dichloro-2-amino pyrimidine (5.0 g, 30.5 mmol) inmethanol (100 mL) was added 25% sodium methoxide (6.59 g, 30.5 mmol).The solution was refluxed for 20 hours, at which time the methanol wasremoved in vacuo. The residue was dissolved in EtOAc (350 mL), washedwith H₂O (100 mL) and with NaCl_((sat.)) (100 mL), dried over Na₂SO₄,filtered and concentrated yielding 4.4 g (90%) of4-chloro-6-methoxypyrimidine-2-ylamine.

To a solution of 4-chloro-6-methoxypyrimidine-2-ylamine (4.4 g, 27.7mmol) in EtOAc (200 mL) and ethanol (150 mL), was addeddiisopropylethylamine (9.6 mL, 55.3 mmol) and 10% palladium on carbon(2.9 g, 2.77 mmol). The heterogeneous solution was stirred under aballoon atmosphere of H₂ for 14 hours, at which time the solution wasfiltered through a Celite pad and the volatiles were removed in vacuo.The residue was dissolved in EtOAc (200 mL), washed with Na₂CO_(3(sat.))(100 mL) and with NaCl_((sat.)) (100 mL), dried over Na₂SO₄, filteredand concentrated yielding 3.1 g (90%) of 4-methoxypyrimidine-2-ylamine.LCMS (m/z): 126 (MH⁺). ¹H NMR (CDCl₃): δ 8.00 (d, J=5.7 Hz, 1H), 6.08(d, J=5.7 Hz, 1H), 4.98 (bs, 2H), 3.84 (s, 3H).

Synthesis of 5-bromo-4-methoxypyrimidine-2-ylamine

To a solution of 4-methoxypyrimidine-2-ylamine (1.84 g, 14.7 mmol) inchloroform (600 mL) was added N-bromosuccinimide (2.62 g, 14.7 mmol).After stirring in the dark for 5 hours, the solution was added to CH₂Cl₂(200 mL) and 1N NaOH (100 mL). Upon mixing, the layers were separatedand the organic layer was washed with NaCl_((sat.)) (100 mL), dried overNa₂SO₄, filtered and concentrated yielding 2.88 g (96%) of5-bromo-4-methoxypyrimidine-2-ylamine. LCMS (m/z): 204/206 (MH⁺). ¹H NMR(CDCl₃): δ 8.10 (s, 1H), 4.93 (bs, 2H), 3.96 (s, 3H).

Synthesis of4-methoxy-5-(4,4,5,5-tetramethyl(1,3,2-dioxaborolan-2-yl))pyrimidine-2-ylamine

To a dry 200-mL flask was added 5-bromo-4-methoxypyrimidine-2-ylamine(2.88 g, 14.1 mmol), potassium acetate (4.16 g, 42.4 mmol),4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane(3.76 g, 14.8 mmol) and dioxane (75 mL). Argon was bubbled through thesolution for 15 minutes, at which time1,1′-bis(diphenylphosphino)ferrocene palladium(II) chloridedichloromethane adduct (0.58 g, 0.71 mmol). The reaction was refluxed ina 115° C. oil bath for 21 hours under argon. After cooling to roomtemperature, the dioxane was removed in vacuo. EtOAc (500 mL) was addedand the resulting slurry was sonicated and filtered. Additional EtOAc(500 mL) was used to wash the solid. The combined organics wereconcentrated and the crude material was purified by SiO₂ chromatography(EtOAc as eluent) yielding 2.4 g of an off white solid. By ¹H NMR thematerial was a 1:1 mixture of boronate ester and4-methoxypyrimidine-2-ylamine. The material was used as is in subsequentSuzuki reactions. LCMS (m/z): 170 (MH⁺ of boronic acid, deriving from insitu product hydrolysis on LC). ¹H NMR (CDCl₃): δ 8.42 (s, 1H), 5.22(bs, 2H), 3.90 (s, 3H), 1.34 (s, 12H).

Method 9 Synthesis of 5-bromo-6-fluoro-2-pyridylamine

To a solution of 6-fluoro-2-pyridylamine (1.0 g, 8.93 mmol) inchloroform (55 mL) was added N-bromosuccinimide (1.59 g, 8.93 mmol). Thesolution was stirred in the dark for 15 hours, at which time it wasadded to CH₂Cl₂ (200 mL) and 1N NaOH (50 mL). Upon mixing, the layerswere separated and the organic layer was washed with NaCl_((sat.)) (50mL), dried over Na₂SO₄, filtered and concentrated. The crude materialwas purified by SiO₂ chromatography (25% EtOAc/hexanes) yielding5-bromo-6-fluoro-2-pyridylamine (386 mg, 22%). LCMS (m/z): 190.9/192.9(MH⁺); ¹H NMR (CDCl₃): δ 7.59 (t, J=8.7 Hz, 1H), 6.25 (dd, J=8.1, 1.2Hz, 1H), 4.58 (bs, 1H).

Synthesis of6-fluoro-5-(4,4,5,5-tetramethyl(1,3,2-dioxaborolan-2-yl))-2-pyridylamine

To a dry 50-mL flask was added 5-bromo-6-fluoro-2-pyridylamine (370 mg,1.93 mmol), potassium acetate (569 mg, 5.8 mmol),4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane(538 mg, 2.12 mmol) and dioxane (15 mL). Argon was bubbled through thesolution for 15 minutes, at which time1,1′-bis(diphenylphosphino)ferrocene palladium(II) chloridedichloromethane adduct (79 mg, 0.09 mmol). The reaction was refluxed ina 115° C. oil bath for 4 hours under argon. After removal of thevolatiles in vacuo, EtOAc (150 mL) was added and the solution was washedwith H₂O (3×40 mL), with NaCl_((sat.)) (300 mL), dried over Na₂SO₄,filtered and concentrated. Purification by SiO₂ chromatography (30%EtOAc/hexanes) yielded boronate ester (161 mg, 35%). LCMS (m/z): 157(MH⁺ of boronic acid, deriving from in situ product hydrolysis on LC) ¹HNMR (CDCl₃): δ 7.86 (t, J=8.4 Hz, 1H), 6.29 (dd, J=8.1, 2.7 Hz, 1H),4.70 (bs, 1H), 1.32 (s, 12H).

Method 10 Synthesis of 5-bromo-4-fluoropyridin-2-amine

N-Bromosuccinimide (126 mg, 0.71 mmol) was added to a solution of4-fluoropyridin-2-amine TFA salt (162 mg, 0.72 mmol) in acetonitrile (4mL) in an aluminum foil-wrapped flask in a darkened hood. The reactionsolution was stirred at room temperature in darkness for 2 hours. Afterevaporation of the solvent, the crude product was purified on a silicagel column eluting with EtOAc to give 5-bromo-4-fluoropyridin-2-amine asan ivory solid (92 mg, 67%). LC/MS (m/z): 190.9/192.9 (MH⁺), R_(t) 1.02minutes.

Synthesis of4-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine

In a sealable Pyrex pressure vessel, a mixture of5-bromo-4-fluoropyridin-2-amine (25 mg, 0.13 mmol),4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane(40 mg, 0.16 mmol), potassium acetate (51 mg, 0.52 mmol) anddichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium(II)-dichloromethaneadduct (16 mg, 0.019 mmol) was suspended in dioxane (1.7 mL) underargon. The pressure vessel was sealed and the reaction mixture wasstirred at 110° C. for 2 hours. After the reaction was complete asjudged by LCMS, the reaction mixture was cooled to room temperature andthe4-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-aminewas used in subsequent reactions without further purification, assuminga quantitative yield (0.13 mmol). LC/MS (m/z): 157.0 (MH⁺ of the boronicacid formed by product hydrolysis on LC), R_(t) 0.34 minutes.

Method 11 Synthesis of 2-amino-5-bromo-isonicotinonitrile

In an aluminum foil-covered flask in a darkened hood,2-amino-isonicotinonitrile TFA salt (125 mg, 0.54 mmol) was dissolved inacetonitrile (3.5 mL). Solid N-bromosuccinimide (89.2 mg, 0.501 mmol)was added to the stirred solution in one portion at RT. The reactionsolution was stirred at room temperature in darkness for 90 minutes.After evaporation of the solvent, the crude material was furtherpurified by silica gel chromatography to give2-amino-5-bromo-isonicotinonitrile (53 mg, 49%). LC/MS (m/z): 197.9(MH⁺), R_(t) 2.92 minutes.

Synthesis of 2-amino-5-boronic ester-isonicotinonitrile

In a glass pressure vessel, a mixture of2-amino-5-bromo-isonicotinonitrile (25 mg, 0.126 mmol),4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane(38 mg, 0.151 mmol) and potassium acetate (49 mg, 0.504 mmol) weresuspended in dioxane (1.8 mL). After purging the mixture with argon for1-2 min, dichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium(II)dichloromethane adduct (16 mg, 0.019 mmol) was added in one portion. Thereaction vessel was sealed and heated at 120° C. with stirring for 2hours. The crude reaction solution was cooled to room temperature andused without further purification assuming a quantitative yield of theboronic ester (0.126 mmol). LC/MS (m/z): 164.0 (MH⁺ of the boronic acidformed by product hydrolysis on LC), R_(t) 0.37 minutes.

Method 12 Synthesis of3-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amineSynthesis of N-allyl-3-fluoropyridin-2-amine

To a preformed bright-yellow complex of Pd(dppf)Cl₂ CH₂Cl₂ (41 mg, 0.05mmol), dppf (83 mg, 0.15 mmol) and NaOt-Bu (1.4 g, 15 mmol) in THF (20mL) was added 2-chloro-3-fluoropyridine (1.32 g, 10 mmol) and allylamine(1.2 mL, 15 mmol). The mixture was sparged with nitrogen and thepressure vessel was capped and sealed. The reaction was heated at 65-70°C. for 16 hours. The cooled reaction was filtered through a plug ofCelite and the pad was washed with EtOAc (30 mL). The solvent wasremoved under reduced pressure to give a brown thick oil. The crudeproduct was purified by silica gel chromatography eluting with 5% MeOHin EtOAc. The product-containing fractions were diluted with EtOAc (100mL) and extracted with 1 M HCl (2×50 mL). The aqueous acidic product waslyophilized to a light brown solid givingN-allyl-3-fluoropyridin-2-amine as an HCl salt (1.6 g, 85%). LC/MS(m/z): 153.1 (MH⁺), R_(t) 0.5 minutes.

Synthesis of 3-fluoropyridin-2-amine

In one portion, 10% Pd/C (1.23 g) was added to a solution ofN-allyl-3-fluoropyridin-2-amine (1.62 g, 7.18 mmol) and BF₃.Et₂O (900uL, 7.18 mmol) in EtOH (20 mL) at RT under nitrogen. After stirring at80° C. for 2 days, the reaction mixture was filtered through a plug ofCelite and the pad was washed with EtOH (20 mL). 6 N HCl was added tothe light yellow filtrate until the solution was acidic. The HCl salt of3-fluoropyridin-2-amine is much less volatile than the free base. Thefiltrate was concentrated under reduced pressure. The salt residue wasdried in vacuo to give 3-fluoropyridin-2-amine as a light yellow glassysolid (1.66 g, quant. yield). LC/MS (m/z): 113.0 (MH⁺), R_(t) 0.41minutes.

Synthesis of 5-bromo-3-fluoropyridin-2-amine and3-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine

Solid NBS (750 mg, 4.2 mmol) was added to a solution of3-fluoropyridin-2-amine HCl salt (1.66 g, 7.18 mmol) in ACN (30 mL) atRT with stirring. The reaction was shielded from light and stirred undernitrogen. After 1 h, an additional amount of NBS (250 mg, 1.4 mmol) wasadded to the reaction. After 1 h, the solvent was removed under reducedpressure and the residue purified by silica gel flash chromatographyeluting with 70% EtOAc/hexane followed by 100% EtOAc to afford5-bromo-3-fluoropyridin-2-amine as a yellow-brown solid (1.26 g, 92%yield). LC/MS (m/z): 191.0/193.0 (MH⁺), R_(t) 1.18 minutes.

The bromide was converted to the pinacolborane ester under conditionsdescribed in Method 1. LC/MS (m/z): 157.0 (MH⁺), R_(t) 0.36 minutes.

Method 13 Synthesis of4-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amineSynthesis of N-allyl-4-fluoropyridin-2-amine

To a preformed red-brown complex of Pd(dppf)Cl₂ (817 mg, 1.0 mmol), dppf(1.66 g, 3.0 mmol) and NaOtBu (2.9 g, 30 mmol) in toluene (30 mL) wasadded 2-chloro-4-fluoropyridine (2.16 g, 20 mmol) and allylamine (1.2mL, 15 mmol). The mixture was sparged with nitrogen and the pressurevessel was capped and sealed. The reaction was heated at 120-125° C. for18 hours. The cooled dark brown reaction was filtered through a plug ofCelite and the pad was washed with EtOAc (60 mL). The solvent was gentlyremoved under reduced pressure to give a brown thick oil which cansublime under vacuum. The crude mixture was acidified with 6 N HCl (10mL) and lyophilized to dryness to give a brown powder as the HCl salt.The crude product was partitioned between EtOAc (100 mL) and sat. NaHCO₃(80 mL) The layers were separated and the aqueous layer was extractedagain with EtOAc (100 mL). The combined organic layers are washed withbrine (100 mL), dried over sodium sulfate, filtered and concentratedunder reduced pressure to give a brown solidN-allyl-4-fluoropyridin-2-amine (690 mg, 25%). LC/MS (m/z): 153.0 (MH⁺),R_(t) 1.13 minutes.

Synthesis of N-allyl-4-fluoropyridin-2-amine

In one portion, 10% Pd/C (552 mg) was added to a solution ofN-allyl-4-fluoropyridin-2-amine (690 mg, 3.07 mmol) and BF₃.Et₂O (0.386mL, 3.07 mmol) in abs. EtOH (12 mL) at RT under nitrogen. After stirringat 80° C. for 24 h, reaction mixture was filtered through a plug ofCelite and the pad was washed with MeOH (100 mL). 6 N HCl (2 mL) wasadded to the dark filtrate until the solution was acidic. The HCl saltof 4-fluoropyridin-2-amine is much less volatile than the free base. Thefiltrate was concentrated under reduced pressure and dried in vacuo. Thecrude product was purified by preparative HPLC to give4-fluoropyridin-2-amine as a brown powder TFA salt (162 mg, 23%). LC/MS(m/z): 113.0 (MH⁺), R_(t) 0.40 minutes.

Synthesis of 5-bromo-4-fluoropyridin-2-amine and4-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine

Solid NBS (78 mg, 0.43 mmol) was added to a solution of3-fluoropyridin-2-amine HCl salt (162 mg, 0.72 mmol) in ACN (4 mL) at RTwith stirring. The reaction was shielded from light and stirred undernitrogen. After 1.5 h, an additional amount of NBS (15 mg, 0.084 mmol)was added to the reaction. Checking the reaction again after 1.5 h, anadditional amount of NBS (15 mg, 0.084 mmol) was added to the reactionuntil the starting material had been consumed by LCMS. After 1 h, thesolvent was removed under reduced pressure and the residue purified bysilica flash chromatography eluting with 50% ethyl acetate/hexane toafford 5-bromo-4-fluoropyridin-2-amine as a ivory solid (92 mg, 68%).LC/MS (m/z): 190.9/192.9 (MH⁺), R_(t) 1.02 minutes.

The bromide was converted to the pinacolborane under conditionsdescribed in Method 1. LC/MS (m/z): 157.0 (MH⁺), R_(t) 0.34 minutes.

Method 14 Synthesis of 2-(5-nitropyridin-2-yloxy)-N,N-dimethylethanamine

Microwave heating. To a solution of 2-(dimethylamino)ethanol (339 mg,3.80 mmol) in DMF (5 mL) was added sodium bis(trimethylsilyl)amide (4.75mL, 1M solution in THF, 4.75 mmol). The mixture was stirred at roomtemperature for 15 min. 2-Chloro-5-nitropyridine (500 mg, 3.16 mmol) wasthen added. The vial was capped and subjected to microwave irradiation(150° C. for 10 minutes). The mixture was diluted with water (250 mL)and EtOAc (250 mL). The two layers were separated, and the aqueous layerextracted two more times with EtOAc. The organic extracts were combined,washed with water and brine, dried over sodium sulfate and evaporated togive the crude material as brown oil. Purification by columnchromatography on silica gel using 5% methanol/methylene chlorideyielded 2-(5-nitropyridin-2-yloxy)-N,N-dimethylethanamine as a lightyellow solid (295 mg, 44%).

Sodium hydride and oil bath heating. To a mixture of sodium hydride (189mg, 4.73 mmol) in anhydrous tetrahydrofuran (2 mL) at 0° C. a solutionof 2-chloro-5-nitropyridine (500 mg, 3.16 mmol) and2-(dimethylamino)ethanol (353 mg, 3.96 mmol) in anhydroustetrahydrofuran (4 mL) was added dropwise. The reaction was warmed toroom temperature and stirred for 16 h. The THF was evaporated, and water(100 mL) and EtOAc (200 mL) were added. The aqueous layer was extractedwith EtOAc (200 mL), and the organic layers combined, washed with brine,dried over sodium sulfate and concentrated to give a brown oil.Purification column chromatography on silica gel using 5%methanol/methylene chloride yielded2-(5-nitropyridin-2-yloxy)-N,N-dimethylethanamine as a light yellowsolid (233=mg, 35%). LC/MS (m/z): 212.2 (MH⁺), R_(t) 1.28 minutes.

Synthesis of 6-(2-(dimethylamino)ethoxy)pyridin-3-amine

2-(5-Nitropyridin-2-yloxy)-N,N-dimethylethanamine (295 mg, 1.40 mmol)was dissolved in 5 mL of methanol and placed under a nitrogenatmosphere. A catalytic amount of 10% palladium on carbon was added anda hydrogen balloon was connected to the reaction flask. The flask wasflushed five times with hydrogen and stirred at room temperature underhydrogen atmosphere for 16 hours. The solid was filtered and washed withmethanol. The filtrate was evaporated under reduced pressure yielding6-[2-(dimethylamino)ethoxy]pyridin-3-amine as a brown oil (250 mg, 99%).LC/MS (m/z): 182.1 (MH⁺), R_(t) 0.36 minutes.

Method 15 Synthesis of 2-(1-methylpiperidin-4-yloxy)-5-nitropyridine

To a mixture of sodium hydride (189 mg, 4.73 mmol) in anhydroustetrahydrofuran (2 mL) at 0° C., a solution of 2-chloro-5-nitropyridine(500 mg, 3.16 mmol) and 1-methylpiperidin-4-ol (455 mg, 3.96 mmol) inanhydrous tetrahydrofuran (4 mL) was added dropwise. The reaction washeated reflux for 16 h. The THF was evaporated and water (100 mL) andEtOAc (200 mL) were added. The aqueous layer was extracted with EtOAc(200 mL). The organic layers were combined, washed with brine, driedover sodium sulfate and concentrated to give a brown oil. Purificationby silica gel column chromatography using 3% methanol/methylene chlorideyielded 2-(1-methylpiperidin-4-yloxy)-5-nitropyridine as a yellow solid,(367 mg, 49%). LC/MS (m/z): 238.0 (MH⁺), R_(t) 1.59 minutes.

Synthesis of 6-(1-methylpiperidin-4-yloxy)pyridin-3-amine

2-(1-Methylpiperidin-4-yloxy)-5-nitropyridine (100 mg, 0.42 mmol) wasdissolved in 5 mL of methanol and placed under a nitrogen atmosphere. Acatalytic amount of 10% palladium on carbon was added and a hydrogenballoon connected to the reaction flask. The flask was flushed fivetimes with hydrogen and stirred at room temperature under hydrogenatmosphere. The solid was filtered and washed with methanol. Thefiltrate was evaporated under reduced pressure to yield6-(1-methylpiperidin-4-yloxy)pyridin-3-amine as a brown solid (85 mg,98%). LC/MS (m/z): 208.2 (MH⁺), R_(t) 0.34 minutes.

Method 16 Synthesis of tert-butyl4-(5-nitropyridin-2-yloxy)piperidine-1-carboxylate

To a solution of tert-butyl 4-hydroxypiperidine-1-carboxylate (1 eq) inDMF was added potassium bis-trimethylsilylamide (1.5 eq, 1M solution intetrahydrofuran). The solution was stirred at room temperature for 10minutes, and 2-chloro-5-nitropyridine (1.2 eq) was added. The reactionmixture was submitted to microwave irradiation for 600 seconds at 145°C. EtOAc and water were added to the reaction and the layers separated.The organic layer was washed with water, brine, dried over sodiumsulfate and evaporated to give brown crude material. Purification bysilica gel column chromatography using 10% EtOAc/hexane afforded theproduct as a light yellow solid. LC/MS (m/z): 324.3 (MH⁺) R_(t) 3.33minutes

Synthesis of 5-nitro-2-(piperidin-4-yloxy)pyridine

Trifluoroacetic acid (5 eq) was added to a solution of tert-butyl4-(5-nitropyridin-2-yloxy)piperidine-1-carboxylate (1 eq) indichloromethane, stirring at room temperature for 1 hour. The solventwas then evaporated, the residue brought to pH=10 with sat. aq. Na₂CO₃solution and extracted with EtOAc. The organic layer was washed withbrine, dried over sodium sulfate and evaporated to afford the product asa light yellow crystalline solid. LC/MS (m/z): 224.3 (MH⁺), R_(t) 1.64minutes

Synthesis of 2-(1-isopropylpiperidin-4-yloxy)-5-nitropyridine

To a solution of 10% acetic acid in methanol was added5-nitro-2-(piperidin-4-yloxy)pyridine (1 eq) and anhydrous acetone (5eq). The solution was stirred at room temperature for 1 hour. Thereaction mixture was cooled to 0° C. on an ice bath and sodiumcyanoborohydride (1.5 eq) was added. The reaction mixture was thenwarmed to room temperature and stirred for 5 hours. The solvent wasevaporated, the residue brought to pH=10 with sodium carbonate andextracted with EtOAc. The organic layer was washed with water, brine,dried over sodium sulfate, and evaporated to give the crude material.Purification by silica gel column chromatography using 2%methanol/dichloromethane afforded the product as a yellow solid. LC/MS(m/z): 266.3 (MH⁺), R_(t) 1.85 minutes.

Synthesis of 6-(1-isopropylpiperidin-4-yloxy)pyridin-3-amine

2-(1-Isopropylpiperidin-4-yloxy)-5-nitropyridine (1 eq) was dissolved inmethanol and laced under a nitrogen atmosphere. A catalytic amount of10% palladium on carbon was added and a hydrogen balloon connected tothe reaction flask. The flask was flushed five times with hydrogen andstirred at room temperature for 4 hours under hydrogen atmosphere. Thesolid was filtered and washed with methanol. The filtrate was evaporatedunder reduced pressure yielding the product as a brown oil. LC/MS (m/z):236.3 (MH⁺), R_(t) 0.38 minutes.

Method 17 Synthesis of 7-methylthio-3-nitroquinoline

To a refluxing mixture of3-[(3-methylthiophenyl)amino]-2-nitroprop-2-enal (2.3 g, 9.6 mmol) andthe HCl salt of 3-methylthiophenylamine (2.7 g, 19.3 mmol) in aceticacid (25 mL) was added thiophenol (0.2 g, 1.9 mmol). After beingrefluxed for 18 h, the mixture was cooled to room temperature and theacetic acid was removed under reduced pressure. To the remaining darkcolored solid EtOAc (50 mL) was added with stirring. Filtration gave ayellow/green solid and a dark filtrate. The product crystallized fromthe EtOAc solution upon standing. Filtration and rinsing with cold EtOAcgave 330 mg crystalline product. The yellow/green solid was washed with3×250 mL portions of dichloromethane. The dichloromethane washes wereconcentrated to give an additional 150 mg of product (23%). LC/MS (m/z):221.1 (MH⁺), R_(t) 2.54 minutes.

Synthesis of 7-(methylsulfonyl)-3-nitroquinoline

To an ice-bath chilled solution of 7-methylthio-3-nitroquinoline (141mg, 0.6 mmol) in dichloromethane (6 mL) was added MCPBA (221 mg, 1.3mmol) in dichloromethane (3 mL). After warming to room temperature, thewhite precipitate formed was filtered and rinsed with an additional 10mL of dichloromethane to yield the pure product (85 mg, 53%). LC/MS(m/z): 252.9 (MH⁺), R_(t) 1.82 minutes.

Synthesis of 7-(methylsulfonyl)-3-quinolylamine

To a suspension of 7-(methylsulfonyl)-3-nitroquinoline (85 mg, 0.4 mmol)in EtOAc (6 mL,) under argon, was added 10% Pd/C (22 mg, 0.04 mmol). AH₂ balloon was connected to the reaction flask, the flask was purgedwith H₂ three times and the reaction mixture was allowed to stir underH₂ atmosphere for 18 h. Unreacted starting material could be seensettling to the bottom of the flask together with the catalyst. Thesolids were removed from the EtOAc solution by filtration. Evaporationof EtOAc under reduced pressure yielded7-(methylsulfonyl)-3-quinolylamine (22 mg, 30%). LC/MS (m/z): 223.0(MH⁺), R_(t) 1.10 minutes.

Method 18 Synthesis of 6-methoxyquinolin-3-amine

A mixture of 6-methoxy-3-nitroquinoline (Magnus, P. et al., J. Am. Chem.Soc. 119, 5591, 1997; 0.17 g, 0.83 mmol) and Pd/C (10%, 80 mg) in EtOAc(15 mL) was hydrogenated with a hydrogen balloon to give6-methoxyquinolin-3-amine in quantitative yield. LC/MS (m/z): 175.1(MH+), R_(t) 1.54 minutes.

Method 19 Synthesis 6-hydroxy-3-nitroquinoline

6-Methoxy-3-nitroquinoline (Magnus, P. et al., J. Am. Chem. Soc. 119,5591, 1997; 100 mg, 0.49 mmol) was dissolved in hydrogen bromidesolution (47% aq, 2.5 mL, 0.2 M), heated and stirred at 120° C. for 16hours. The reaction mixture was cooled to room temperature, neutralizedwith 6N NaOH, then extracted with EtOAc (150 mL). The organic layer wasdried over Na₂SO₄ and purified by flash chromatography (SiO₂, 40-50%EtOAc/hexanes), obtaining 73 mg (78%) of 6-hydroxy-3-nitroquinoline.LC/MS (m/z): 190.9 (MH⁺), R_(t) 1.97 minutes.

Synthesis of 3-nitro-6-(2-(pyrrolidin-1-yl)ethoxy)quinoline

6-Hydroxy-3-nitroquinoline (148 mg, 0.78 mmol) was dissolved in THF (18mL). 2-(Pyrrolidin-1-yl)ethanol (0.091 mL, 0.78 mmol) andtriphenylphosphine (306 mg, 1.17 mmol) were added. Lastly, diethylazodicarboxylate (0.184 mL, 1.17 mmol) was added and the reactionmixture was allowed to stir at room temperature for 2 hours. The solventwas then concentrated in vacuo and the residue was purified by flashchromatography (SiO₂) to yield 134 mg (60%) of3-nitro-6-(2-(pyrrolidin-1-yl)ethoxy)quinoline. LC/MS (m/z): 288.1(MH⁺), R_(t) 1.80 minutes.

Synthesis of 3-amino-6-(2-(pyrrolidin-1-yl)ethoxy)quinoline

3-Nitro-6-(2-(pyrrolidin-1-yl)ethoxy)quinoline (134 mg, 0.46 mmol) wasdissolved in EtOAc (10 mL) and the solution was sparged with N₂ forseveral minutes. Triethylamine (0.065 mL, 0.46 mmol) was then addedfollowed by a catalytic amount of 10% Pd/C. Sparging with N₂ wasrepeated after each addition. A balloon of H₂ was connected to thereaction flask and the reaction mixture was stirred at room temperatureunder H₂ atmosphere for 48 hours. The mixture was then filtered througha Celite pad and concentrated to obtain crude3-amino-6-(2-(pyrrolidin-1-yl)ethoxy)quinoline, which was used as is inthe next reaction. LC/MS (m/z): 258.1 (MH⁺), R_(t) 0.33 minutes.

Method 20 Synthesis of 5-methoxy-3-nitro-quinoline

Synthesis of 3-(3-methoxy-phenylamino)-2-nitro-propenal

To the HCl salt of 3-methoxy-phenylamine (4.6 g, 28.9 mmol) in 1N HCl(300 mL) was added a solution of 2-nitro-malonaldehyde (2.7 g, 19.3mmol) in 150 mL water. After 30 min, the precipitate was filtered andrinsed with 0.1 N HCl. Air-drying in a Büchner funnel for 18 h gave 3.36g (78%) of a light yellow/green powder. LC/MS (m/z): 245.1 (MH⁺+Na),R_(t) 2.21 minutes.

Synthesis of 5-methoxy-3-nitroquinoline and 7-methoxy-3-nitroquinoline

To the HCl salt of 3-methoxy-phenylamine (4.7 g, 29.7 mmol) in 30 mLacetic acid was added 3-(3-methoxy-phenylamino)-2-nitro-propenal (3.3 g,14.9 mmol). The reaction mixture was heated to reflux, and thiophenol(0.3 mL, 2.98 mmol) was added. After 22 h, the reaction mixture wascooled to room temperature and the solvent was removed in vacuo.Addition of 70 mL EtOAc and filtration gave solid byproduct,7-methoxy-3-nitro-quinoline, and a filtrate, which contained impure5-methoxy-3-nitro-quinoline. The filtrate was loaded on to silica columnand eluted from 5% to 25% EtOAc in hexanes at 85 mL/min for 30 minutes.The product-enriched fractions were concentrated and taken on to thenext step as a mixture of 5- and 7-methoxy substituted3-nitroquinolines. LC/MS (m/z): 205.1 (MH⁺), R_(t) 2.26 minutes.

Synthesis of 5-methoxyquinolin-3-amine

A mixture of 5- and 7-methoxy substituted 3-nitroquinolines (780 mg,3.82 mmol) was dissolved in EtOAc (75 mL) and the reaction mixturesparged with N₂ for several minutes. 10% Pd/C (54 mg) was then added anda H₂ balloon was connected to the reaction flask. The reaction mixturewas sparged with H₂ and stirred at room temperature under H₂ atmosphereovernight. Solvent removal in vacuo and purification by columnchromatography on silica gel (100% EtOAc) afforded the two separatedisomers 5-methoxyquinolin-3-amine and 7-methoxyquinolin-3-amine. Thedesired product 5-methoxyquinolin-3-amine (80 mg, 12%) was obtained as ayellow powder. The structure was assigned by ¹H NMR (CD₃OD): δ 8.40 (d,1H), 7.69 (d, 1H), 7.40 (d, 1H), 7.30 (t, 1H), 6.85 (d, 1H). LC/MS(desired isomer) (m/z): 175.0 (MH⁺), R_(t) 1.54 minutes; LC/MS(undesired isomer) (m/z): 175.0 (MH⁺), R_(t) 1.53 minutes.

Method 21 Synthesis of 2-(methylsulfonyl)pyridin-4-amine

Synthesis of 2-(methylthio)pyridin-4-amine

Sodium thiomethoxide (140 mg, 1.98 mmol) was added to a solution of2-chloropyridin-4-amine (150 mg, 1.17 mmol) in NMP (0.65 mL) in apressure vessel. The vessel was sealed and heated in a microwave to 200°C. for 800 sec. Purification by silica flash chromatography eluting with8% MeOH/DCM afforded 2-(methylthio)pyridin-4-amine (435 mg, 50% yield).LC/MS (m/z): 140.9 (MH⁺), R_(t) 0.59 minutes.

Solid MCPBA (780 mg, 2-3 mmol) was slowly added in small portions to asolution of 2-(methylthio)pyridin-4-amine (435 mg, 1.17 mmol) in THF (7mL) at RT, with stirring. The reaction was followed by LCMS as thestarting material was consumed by titrating with MCPBA. Silica was addedto the reaction mixture, which was then concentrated to dryness underreduced pressure. Silica supported crude was purified by silica flashchromatography, eluting with 5% MeOH/DCM, to afford2-(methylsulfonyl)pyridin-4-amine (220 mg, quant. yield). LC/MS (m/z):173.0 (MH⁺), R_(t) 0.34 minutes.

Method 22 Synthesis of 2-morpholinopyrimidine-4,6-diol

Sodium (17.25 g, 150 mmol) was cut into small pieces and slowly added toEtOH (500 mL) in a 1-L round bottom flask under N₂ and cooled withwater. After all the sodium was dissolved, morpholinoformamidinehydrobromide (52.5 g, 50 mmol) and diethyl malonate (40 g, 50 mmol) wereadded. The mixture was heated to reflux for three hours. The reactionmixture was cooled to room temperature, and the ethanol was removed invacuo. Aqueous HCl (1N, 800 mL) was added, to the white solid, at roomtemperature. The solid initially dissolved, giving a clear solution,then the product crashed out as a white solid. After 1 h at roomtemperature, the solid was filtered, washed with water (3×), dried (airand then over P₂O₅) to give 2-morpholinopyrimidine-4,6-diol (42.5 g,86%). LC/MS (m/z): 198.1 (MH⁺), R_(t) 0.51 minutes.

Synthesis of 4,6-dichloro-2-morpholinopyrimidine

A mixture of 2-morpholinopyrimidine-4,6-diol (30 g, 0.15 mol) and POCl₃(150 mL, 1.61 mol) was heated at 120° C. for 16 h, then cooled to RT.Excess POCl₃ was removed to give a semi-solid. The solid was graduallytransferred to a stirring solution of water (700 mL) and EtOH (100 mL)occasionally cooled with water. White solid formed and was subsequentlyfiltered, washed with water, 10% EtOH in water, and dried over P₂O₅ togive 4,6-dichloro-2-morpholinopyrimidine (17.82 g, 50%). LC/MS (m/z):233.9 (MH⁺), R_(t) 2.95 minutes.

Method 23 Synthesis of 4,6-dichloro-5-methyl-2-morpholinopyrimidine

4,6-Dichloro-5-methyl-2-morpholinopyrimidine was prepared by similarprocedure as 4,6-dichloro-2-morpholinopyrimidine (in Method 22) usingdimethyl 2-methylmalonate in place of diethyl malonate. LC/MS (m/z):248.1 (MH⁺).

Method 24 Synthesis of 4,6-dichloro-5-ethyl-2-morpholinopyrimidine

4,6-Dichloro-5-ethyl-2-morpholinopyrimidine was prepared by similarprocedure as 4,6-dichloro-2-morpholinopyrimidine (in Method 22) usingdimethyl 2-ethylmalonate in place of diethyl malonate. LC/MS (m/z):262.1 (MH⁺), R_(t) 3.59 minutes.

Method 25 Synthesis of 5-fluoro-2-morpholinopyrimidine-4,6-diol

Sodium hydride (60% in oil, 3.9 g, 96.5 mmol) was washed with hexanes ina round bottom flask under argon and cooled with an ice water bath. EtOH(100 mL) was slowly added. The resulting mixture was warmed to RT andstirred for 30 minutes. To the base mixture, diethyl 2-fluoromalonate(5.7 g, 32.2 mmol) was added, followed by morpholinoformamidinehydrobromide (6.8 g, 32.2 mmol). The mixture was heated to 90-95° C.with stirring under argon. After 12 hours, the reaction was cooled toroom temperature and the EtOH was removed in vacuo. The resulting whitesolid was dissolved in water (25 mL) and acidified with conc. HCl topH=3-4. A white precipitate formed which was collected on a Büchnerfilter, washed with water (2×50 mL), air dried on the filter, and driedin vacuo to give 5-fluoro-2-morpholinopyrimidine-4,6-diol (0.87 g, 12%).LC/MS (m/z): 216.0 (MH⁺), R_(t) 0.63 minutes.

Synthesis of 4-(4,6-dichloro-5-fluoropyrimidin-2-yl)morpholine

A mixture of 5-fluoro-2-morpholinopyrimidine-4,6-diol (0.87 g, 4.0 mmol)and POCl₃ (10 mL) was heated at 120° C. for 16 hours, then cooled to RT.Excess of POCl₃ was removed under reduced pressure to give a semi-solidwhich was dried further in vacuo. After 12 h of vacuum drying, the solidwas diluted in EtOAc (150 mL) and washed with sat. NaHCO₃ (60 mL). Asolid formed during the wash and was discarded with the aqueous layer.The organic layer was washed again with sat. NaHCO₃ (2×30 mL), brine (30mL), dried with Na₂SO₄, filtered and evaporated under reduced pressureto give a crude product. The product was purified by flashchromatography eluting with 25% EtOAc/hexane to give4-(4,6-dichloro-5-fluoropyrimidin-2-yl)morpholine (418 mg, 42%). LC/MS(m/z): 251.9 (MH⁺), R_(t) 3.22 minutes.

Method 26 Synthesis of 2,4,6-tribromopyrimidine

To a mixture of pyrimidine-2,4,6(1H,3H,5H)-trione (2.66 g, 20.87 mmol)and POBr₃ (25 g, 87.2 mmol) in toluene (35 mL) in a 200 mL flask,N,N-dimethylaniline (4.52 mL, 35.7 mmol) was added. The brick-red slurrywas heated to reflux for 3 hours. During the process a biphasic solutionformed with a red gum at the bottom of the flask and a clear yellowliquid above. The reaction mixture was cooled to room temperature andthe yellow organic layer decanted off. The red gum was rinsed once withEtOAc. The combined organic extracts were washed with saturated NaHCO₃(3×, or until CO₂ evolution ceased), H₂O (3×), brine (2×) and dried overNa₂SO₄. The solution was then concentrated and dried under high vacuumto yield 2,4,6-tribromopyrimidine (5.40 g, 82%), which was used withoutfurther purification. LC/MS (m/z): 316.8/318.7 (MH⁺), R_(t) 2.78minutes.

Method 27

A solution of morpholine (100 g; 1.15 moles; 5.3 equivalents) in THF(450 mL) was cooled with an ice bath. A solution of2,4,6-trichloropyrimidine (39.9 g; 217 mmoles; 1.0 equivalents) in THF(100 mL) was added over a period of 30 minutes. A copious whiteprecipitate formed upon addition of 2,4,6-trichloropyrimidine and thereaction mixture rapidly thickened. The mixture was allowed to warm toambient temperature and mechanically stirred for 64 hours (heating thereaction mixture at reflux following the addition of2,4,6-trichloropyrimidine leads to complete reaction in 60 min. Theratio of a to b was unchanged). The mixture was then filtered and thefilter cake washed with additional THF (2×100 mL). The filtrate wasconcentrated on the rotavap. Water (600 mL) was added and the resultingslurry was stirred for 30 minutes. The solids were isolated byfiltration, washed with additional water (2×100 mL) and dried overnightunder vacuum. Yield a+b: 61.3 g (99%). Product was 87% a by hplc areapercent; remainder is b.

31 g of the crude solid was dissolved in 200 mL of CH₂Cl₂ and applied to600 g of dry silica in a fritted glass funnel. The silica was elutedwith 1:1 hexane:EtOAc and 300 mL fractions were collected. TLC analysisshows a to be present in fractions 1-7 and4,6-dimorpholino-2-chloropyrimidine in fractions 6-10. Fractions 1-5were pooled and concentrated to provide a white solid. Yield: 28.2 g(Product was 98% a by hplc area percent).

Method 28 Synthesis of 4-(1-isopropylpiperidin-4-yloxy)aniline Synthesisof T-butyl 4-(2-methoxy-4-nitrophenoxy)piperidine-1-carboxylate

To a mixture of triphenylphosphine (3.10 g, 11.8 mmol) anddiethylazodicarboxylate (2.06 g, 11.8 mmol) under N₂ in THF (40 mL) wasadded t-butyl 4-hydroxypiperidine-1-carboxylate (2.00 g, 9.94 mmol).After stirring 10 min, 2-methoxy-4-nitrophenol (1.00 g, 5.91 mmol) wasadded. The reaction was stirred for 16 h and the solvent was evaporatedunder reduced pressure to give orange oil. The crude product waspurified by column chromatography on silica gel using 25% EtOAc/hexaneyielding t-butyl 4-(2-methoxy-4-nitrophenoxy)piperidine-1-carboxylate asa beige solid (1.70 g, 82%). LC/MS (m/z): 353.2 (MH⁺), R_(t) 3.23minutes

Synthesis of 4-(2-methoxy-4-nitrophenoxy)piperidine

Trifluoroacetic acid (5 eq) was added to a solution of tert-butyl4-(2-methoxy-4-nitrophenoxy)piperidine-1-carboxylate (200 mg, 0.57 mmol,1 eq) in dichloromethane, stirring at room temperature for 1 hour. Thesolvent was then evaporated, the residue was brought to pH 10 with sat.aq. Na2CO3 solution and extracted with EtOAc. The organic layer waswashed with brine, dried over sodium sulfate and evaporated to affordthe product 4-(2-methoxy-4-nitrophenoxy)piperidine as a light yellowsolid (137.3 mg, 96%). LC/MS (m/z): 253.2 (MH+), Rt 1.81 minutes.

Synthesis of 1-isopropyl-4-(2-methoxy-4-nitrophenoxy)piperidine

To a solution of 10% acetic acid in methanol was added4-(2-methoxy-4-nitrophenoxy)piperidine (148 mg, 0.59 mmol, 1 eq),anhydrous acetone (5 eq), and sodium cyanoborohydride (1.5 eq). Thesolution was stirred at room temperature for 24 h. Additional anhydrousacetone (5 eq) and sodium cyanoborohydride (1.5 eq) were added and thereaction was stirred for 24 h. The solvent was evaporated, the residuewas brought to pH 10 with aqueous sodium carbonate and extracted withEtOAc. The organic layer was washed with water, brine, dried withmagnesium sulfate and evaporated to afford1-isopropyl-4-(2-methoxy-4-nitrophenoxy)piperidine as a yellow oil (163mg, 97%). LC/MS (m/z): 295.2 (MH⁺), R_(t) 1.96 minutes.

Synthesis of 4-(1-isopropylpiperidin-4-yloxy)-3-methoxyaniline

1-isopropyl-4-(2-methoxy-4-nitrophenoxy)piperidine (167 mg, 0.57 mmol)was dissolved in methanol (20 mL) and placed under a nitrogenatmosphere. A catalytic amount of 20% palladium hydroxide on carbon wasadded and a hydrogen balloon was connected to the reaction flask. Theflask was flushed five times with hydrogen and stirred at roomtemperature under hydrogen atmosphere for 16 hours. The reaction mixturewas filtered and washed with methanol. The filtrate was evaporated underreduced pressure. Acetonitrile (10 mL) was added to the residue, swirledfor 10 min, and decanted away from white film. The acetonitrile layerwas evaporated under reduced pressure yielding4-(1-isopropylpiperidin-4-yloxy)aniline as a brown oil (131 mg, 87%).LC/MS (m/z): 265.2 (MH⁺), R_(t) 0.33 minutes.

Method 29 Synthesis of4-(1-isopropylpiperidin-4-yloxy)-3-methoxyaniline; Synthesis ofTert-butyl 4-(2-methoxy-4-nitrophenoxy)piperidine-1-carboxylate

To a mixture under N₂ of triphenylphosphine (3.10 g, 11.825 mmol) anddiethylazodicarboxylate (2.06 g, 11.825 mmol) in THF (40 mL) was addedtert-butyl 4-hydroxypiperidine-1-carboxylate (2.00 g, 9.937 mmol). Afterstirring 10 min, 2-methoxy-4-nitrophenol (1.00 g, 5.912 mmol) was added.The reaction stirred 16 h and evaporated under reduced pressure to givean orange oil. The crude product was purified by column chromatographyon silica gel using 25% EtOAc/hexane yielding tert-butyl4-(2-methoxy-4-nitrophenoxy)piperidine-1-carboxylate as a beige solid(1.70 g, 82%). LC/MS (m/z): 353.2 (MH⁺), R_(t) 3.23 minutes

Synthesis of 4-(2-methoxy-4-nitrophenoxy)piperidine

Trifluoroacetic acid (5 eq) was added to a solution of tert-butyl4-(2-methoxy-4-nitrophenoxy)piperidine-1-carboxylate (200 mg, 0.5676mmol, 1 eq) in dichloromethane, stirring at room temperature for 1 hour.The solvent was then evaporated, the residue brought to pH=10 with sat.aq. Na₂CO₃ solution and extracted with EtOAc. The organic layer waswashed with brine, dried over sodium sulfate and evaporated to affordthe product 4-(2-methoxy-4-nitrophenoxy)piperidine as a light yellowsolid (137.3 mg, 96%). LC/MS (m/z): 253.2 (MH⁺), R_(t) 1.81 minutes.

Synthesis of 1-isopropyl-4-(2-methoxy-4-nitrophenoxy)piperidine

To a solution of 10% acetic acid in methanol was added4-(2-methoxy-4-nitrophenoxy)piperidine (148 mg, 0.59 mmol, 1 eq),anhydrous acetone (5 eq), and sodium cyanoborohydride (1.5 eq). Thesolution was stirred at room temperature for 24 h. Reaction is 85%complete. Charged additional anhydrous acetone (5 eq) and sodiumcyanoborohydride (1.5 eq) and stirred for 24 h. The solvent wasevaporated, the residue brought to pH=10 with sodium carbonate andextracted with EtOAc. The organic layer was washed with water, brine,dried with magnesium sulfate and evaporated to afford1-isopropyl-4-(2-methoxy-4-nitrophenoxy)piperidine as a yellow oil (163mg, 97%). LC/MS (m/z): 295.2 (MH⁺), R_(t) 1.96 minutes.

Synthesis of 4-(1-isopropylpiperidin-4-yloxy)-3-methoxyaniline

1-isopropyl-4-(2-methoxy-4-nitrophenoxy)piperidine (167 mg, 0.57 mmol)was dissolved in 20 mL of methanol and placed under a nitrogenatmosphere. A catalytic amount of 20% palladium hydroxide on carbon wasadded and a hydrogen balloon was connected to the reaction flask. Theflask was flushed five times with hydrogen and stirred at roomtemperature under hydrogen atmosphere for 16 hours. The reaction mixturewas filtered and washed with methanol. The filtrate was evaporated underreduced pressure. Acetonitrile (10 mL) was added to the residue, swirledfor 10 min, and decanted away from white film. The acetonitrile layerwas evaporated under reduced pressure yielding4-(1-isopropylpiperidin-4-yloxy)aniline as a brown oil (131 mg, 87%).LC/MS (m/z): 265.2 (MH⁺), R_(t) 0.33 minutes.

Method 30 Synthesis ofN-(6-chloro-2-morpholinopyrimidin-4-yl)-4-phenylthiazol-2-amine

To a solution of the 4-phenylthiazol-2-amine (374 mg, 2.1 mmol) in 10 mLof N,N-dimethylacetamide was added sodium hydride (50 mg, 2.1 mmol) atroom temperature. After the mixture was stirred at that temperature for10 minutes, the dichloride (470 mg, 2.0 mmol) was added to the reactionmixture. After being stirred at room temperature for 1 hour, additionalsodium hydride (50 mg, 2.1 mmol) was added to the reaction mixture. Themixture was stirred for 1 hour and quenched with 5 mL of aq. ammoniumchloride. The resulting mixture was extracted with ethyl acetate (2×10mL). The combined organic layers were washed with water (10 mL), brine(10 mL), then dried over MgSO4, filtered, and evaporated under reducedpressure to give crude product, which was purified by silica gel columneluted with ethyl acetate and hexane to giveN-(6-chloro-2-morpholinopyrimidin-4-yl)-4-phenylthiazol-2-amine. LC/MS(m/z): 374 and 376 (MH+), Rt 3.40 minutes.

Example 1 Preparation ofN-(6-(2-aminopyrimidin-5-yl)-2-morpholinopyrimidin-4-yl)quinolin-3-amine

4,6-Dichloro-2-morpholinopyrimidine (prepared as in Method 22; 3.0 g,12.9 mmol) and5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidin-2-amine (3.43g, 15.5 mmol) were dissolved in DME (130 mL). Aqueous Na₂CO₃ (2 M, 32mL, 64 mmol) was then added and the reaction mixture was sparged with N₂for several minutes. Pd(OAc)₂ (145 mg, 0.65 mmol) and PPh₃ (339 mg, 1.29mmol) were then added and the reaction mixture was heated at 95° C. for1 h. The reaction mixture was allowed to cool to room temperature, thesolution was decanted away from the solid residue and concentrated. Thesolid thus formed was separated from the water phase. The waterextracted with EtOAc and this organic layer was combined with theprecipitate. Removal of the solvent in vacuo gave a solid residue whichwas triturated with about 20 mL of EtOAc, filtered and evaporated underreduced pressure to give the desired product. Additional product wasobtained by concentrating the mother liquor and purifying the solidcrash out by trituration with EtOAc. The two crops were combinedobtaining 1.98 g (52%) of the desired product. LC/MS (m/z): 293.1 (MH⁺),R_(t) 1.92 minutes

N-(6-(2-aminopyrimidin-5-yl)-2-morpholinopyrimidin-4-yl)quinolin-3-amine

Pd(OAc)₂, BINAP, cesium carbonate, THF (0.8 mL) were mixed with5-(6-chloro-2-morpholinopyrimidin-4-yl)pyrimidin-2-amine (1 eq) andquinolin-3-amine (2 eq). The mixture was heated under microwaveirradiation for 10 minutes at 110° C. The solution was filtered andconcentrated under reduced pressure. LC/MS (m/z): 401.4 (MH⁺).

Example 2 Preparation ofN-(6-(6-aminopyridin-3-yl)-2-morpholinopyrimidin-4-yl)quinolin-3-amine5-(6-Chloro-2-morpholin-4-yl-pyrimidin-4-yl)pyridin-2-ylamine

THF (130 mL) and aq. Na₂CO₃ (2M, 40 mL, 80 mmol) were added to a glasspressure vessel containing 4,6-dichloro-2-morpholinopyrimidine (preparedas in Method 22; 4.5 g, 19.2 mmol) and5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine (4.7 g,21.2 mmol). The resulting mixture was stirred and sparged with argon for1-2 minutes. The catalyst,dichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium (II)dichloromethane adduct (1.26 g, 1.54 mmol), was then added in oneportion. After sealing the reaction vessel, the reaction was heated at85° C. for 1 hour with stirring. Upon cooling to RT, the THF was removedunder reduced pressure to leave a viscous residue. EtOAc (450 mL) andwater (50 mL) were added. After vigorously stirring for 1-2 minutes, thesolids were filtered off and washed with EtOAc (100 mL). The organiclayer was separated and the aqueous layer was extracted with EtOAc (100mL). The combined organic layers were washed with saturated NaClsolution (1×50 mL), dried with Na₂SO₄, filtered and evaporated underreduced pressure. The crude material was further purified by silica gelchromatography to give5-(6-chloro-2-morpholin-4-yl-pyrimidin-4-yl)-pyridin-2-ylamine (2.48 g,44%). LC/MS (m/z): 292.1 (MH⁺), R_(t) 2.06 minutes.

[6-(6-amino-pyridin-3-yl)-2-morpholin-4-yl-pyrimidin-4-yl]-(6-methoxy-pyridin-3-yl)-amine

In a glass pressure vessel, Pd(OAc)₂ (2.0 mg, 0.0082 mmol), BINAP (6.4mg, 0.0102 mmol), cesium carbonate (20.0 mg, 0.0615 mmol) and THF (0.8mL) were mixed and stirred at room temperature for 1-3 minutes. To theresulting mixture was added5-(6-chloro-2-morpholin-4-yl-pyrimidin-4-yl)-pyridin-2-ylamine (12.0 mg,0.041 mmol) followed by 6-methoxypyridin-3-ylamine (10.2 mg, 0.082mmol). The glass pressure vessel was sealed and stirred at 95° C. for 90minutes. The reaction mixture was filtered and concentrated underreduced pressure. The product was purified by preparative reverse phaseHPLC to give[6-(6-amino-pyridin-3-yl)-2-morpholin-4-yl-pyrimidin-4-yl]-(6-methoxy-pyridin-3-yl)-amine(5.0 mg, 32%). LC/MS (m/z): 380.1 (MH⁺), R_(t) 1.82 minutes.

Example 3 Preparation of5-(6-[2-(methylsulfonamide)pyridin]-3-yl)-2-morpholino-pyrimidin-4-yl)pyridin-2-amine5-[6-(2-fluoro-pyridin-3-yl)-2-morpholin-4-yl-pyrimidin-4-yl]-pyridin-2-ylamine

To a solution of5-(6-chloro-2-morpholin-4-yl-pyrimidin-4-yl)pyridin-2-ylamine, preparedas in Example 2, (252 mg, 0.87 mmol) and2-fluoro-3-(4,4,5,5-tetramethyl-[1,3,2]dioxaborolan-2-yl)pyridine (183mg, 1.30 mmol) in DME (4 mL) was added an aqueous solution of Na₂CO₃ (2M, 1 mL), followed bydichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium(II)-dichloromethane(71 mg, 0.087 mmol). The mixture was heated in a microwave for 20 min at120° C. The aqueous phase was separated from DME, and extracted withEtOAc. The combined organic phases were washed with brine, dried,filtered, and concentrated to give crude desired product which wascarried on to the next step without further purification. LC/MS (m/z):353.3 (MH⁺), 1.84 minutes.

N-{3-[6-(6-Amino-pyridin-3-yl)-2-morpholin-4-yl-pyrimidin-4-yl]pyridin-2-yl}-methanesulfonamide

To a solution of5-[6-(2-fluoro-pyridin-3-yl)-2-morpholin-4-yl-pyrimidin-4-yl]-pyridin-2-ylamine(200 mg, 0.57 mmol) and methanesulfonamide (216 mg, 2.3 mmol) in NMP (8mL) was added Cs₂CO₃ (372 mg, 1.1 mmol). The solution was heated at 125°C. for 4 hours. The reaction mixture was cooled to room temperature,filtered and purified by reverse phase preparatory HPLC to give thetitle compound. LC/MS (m/z): 428.3 (MH⁺), R_(t) 1.80 minutes.

Example 4 Preparation ofN-(6-(6-amino-4-fluoropyridin-3-yl)-2-morpholinopyrimidin-4-yl)quinolin-3-amineN-(6-bromo-2-morpholinopyrimidin-4-yl)quinolin-3-amine

To a solution of 2,4,6-tribromopyrimidine (5.40 g, 17.2 mmol) inacetonitrile (60 mL) was added quinolin-3-amine, followed by DIEA (8.99mL, 51.6 mmol). The reaction mixture was heated to 45° C. overnight.Morpholine (1.50 mL, 17.2 mmol) was then added, and the reaction mixturecontinued heating for 4 h. The reaction mixture was then cooled to roomtemperature, concentrated and dissolved in EtOAc (about 500 mL), theorganic solution was washed with saturated NaHCO₃ (3×), H₂O (2×), brine(1×) and dried over Na₂SO₄. The solution was then evaporated in thepresence of silica gel and purified by column chromatography (SiO₂,15-25% EtOAc/Hexanes) to yieldN-(6-bromo-2-morpholinopyrimidin-4-yl)quinolin-3-amine. LC/MS (m/z):386.1 (MH⁺).

N-(6-(6-amino-4-fluoropyridin-3-yl)-2-morpholinopyrimidin-4-yl)quinolin-3-amine

To a solution of4-fluoro-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine(prepared as shown in Method 10) in dioxane (1.7 mL, 0.13 mmol),N-(6-bromo-2-morpholinopyrimidin-yl)quinolin-3-amine (20 mg, 0.052mmol),dichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium(II)-dichloromethaneadduct (11 mg, 0.013 mmol) and 2 M aqueous sodium carbonate solution(0.6 mL, 1.2 mmol) were added under argon. The pressure vessel wassealed and the reaction mixture was heated in a microwave reactor at120° C. for 15 minutes. The crude product was partitioned between EtOAc(30 mL) and saturated sodium bicarbonate (10 mL). The organic layer wasseparated, dried over sodium sulfate, filtered and concentrated underreduced pressure. The product was purified by reverse phase preparativeHPLC to giveN-(6-(6-amino-4-fluoropyridin-3-yl)-2-morpholinopyrimidin-4-yl)quinolin-3-amineas yellow powder (14 mg, 26%). LC/MS (m/z): 418.0 (MH⁺), R_(t) 2.31minutes.

Example 5 Preparation of2-amino-5-[2-morpholin-4-yl-6-(quinolin-3-ylamino)-pyrimidin-4-yl]-isonicotinonitrile

To the crude2-amino-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridine-4-carbonitrile(prepared as in Method 11) (25 mg, 0.13 mmol) solution in dioxane (1.8mL) in a pressure vessel, was added(6-bromo-2-morpholin-4-yl-pyrimidin-4-yl)-quinolin-3-yl-amine (19.4 mg,0.05 mmol) and aq. Na₂CO₃ (2 M, 0.6 mL, 1.2 mmol). After purging thereaction mixture with argon,dichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium(II)dichloromethane adduct (10.3 mg, 0.01 mmol) was added in one portion.The pressure vessel was sealed and the mixture was heated in a microwaveat 120° C. for 900 seconds. The crude mixture was filtered andconcentrated under reduced pressure. The crude product was purified byreverse phase preparative HPLC to give2-amino-5-[2-morpholin-4-yl-6-(quinolin-3-ylamino)-pyrimidin-4-yl]-isonicotinonitrile(4.5 mg, 20%). LC/MS (m/z): 425.0 (MH⁺), R_(t) 2.03 minutes.

Example 6 Preparation ofN⁶-methyl-2-morpholino-N⁶-(tetrahydro-2H-pyran-4-yl)-4,5′-bipyrimidine-2′,6-diamine

N-Methyltetrahydro-2H-pyran-4-amine

Tetrahydro-2H-pyran-4-amine (90 mg, 0.9 mmol) was added to a solution offormaldehyde (37% solution in water, 0.091 mL, 1.13 mmol) and aceticacid (0.162 mL) in ACN (0.8 mL). After stirring for 5 minutes, Na(CN)BH₃(60 mg, 1.13 mmol) was added in one portion at RT. After 1 hour, excessCs₂CO₃ was added to the reaction until made alkaline. After stirring for15 minutes, the reaction was filtered to remove solids and the solventevaporated under reduced pressure. The crude product,N-methyltetrahydro-2H-pyran-4-amine, was used for the followingdisplacement without further purification. LC/MS (m/z): 116.1 (MH⁺),R_(t) 0.34 minutes.

6-chloro-N-methyl-2-morpholino-N-(tetrahydro-2H-pyran-4-yl)pyrimidin-4-amine

The crude N-methyltetrahydro-2H-pyran-4-amine (104 mg, 0.9 mmol) wasdissolved in NMP (0.8 mL). To the solution, Cs₂CO₃ (366 mg, 1.13 mmol)and 4-(4,6-dichloropyrimidin-2-yl)morpholine (prepared as in Method 22)(80 mg, 0.34 mmol) were added at room temperature. The reaction mixturewas heated to 95° C. After 90 minutes, the reaction mixture was cooledto room temperature, filtered and purified by reverse phase preparativeHPLC yielding 24 mg (23%) of pure6-chloro-N-methyl-2-morpholino-N-(tetrahydro-2H-pyran-4-yl)pyrimidin-4-amine.LC/MS (m/z): 313.2 (MH⁺), R_(t) 2.61 minutes.

N⁶-methyl-2-morpholino-N⁶-(tetrahydro-2H-pyran-4-yl)-4,5′-bipyrimidine-2′,6-diamine

To an argon flushed mixture of5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidin-2-amine (42 mg,0.19 mmol),6-chloro-N-methyl-2-morpholino-N-(tetrahydro-2H-pyran-4-yl)pyrimidin-4-amine(12 mg, 0.038 mmol) in THF (0.8 mL), and aq. Na₂CO₃ (2M, 0.27 mL) in apressure vessel, was addeddichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium (II)dichloromethane adduct (8 mg, 0.0095 mmol) in one portion. The pressurevessel was sealed and the mixture was heated in a microwave at 120° C.for 600 seconds. The crude mixture was filtered, concentrated underreduced pressure and purified by reverse phase preparative HPLC to giveN⁶-methyl-2-morpholino-N⁶-(tetrahydro-2H-pyran-4-yl)-4,5′-bipyrimidine-2′,6-diamine(4.6 mg, 32%). LC/MS (m/z): 372.2 (MH⁺), R_(t) 1.76 minutes.

Example 7 Preparation ofN-(6-(2-aminopyrimidin-5-yl)-2-morpholinopyrimidin-4-yl)-5-methoxyquinolin-3-amine

The desired compound was prepared as described in Example 2: Pd(OAc)₂,BINAP, cesium carbonate, THF (0.8 mL) were mixed with5-(6-chloro-2-morpholinopyrimidin-4-yl)pyrimidin-2-amine (1 eq.) and5-methoxyquinolin-3-amine (2 eq), which was prepared as shown in Method20. The mixture was heated under microwave irradiation for 10 minutes at110° C. The solution was filtered and concentrated under reducedpressure. The crude product was purified by preparative reverse phaseHPLC. LC/MS (m/z): 431.2 (MH⁺), R_(t) 2.03 minutes.

Example 8 Preparation of5-(2-morpholino-6-(pyridin-3-yloxy)pyrimidin-4-yl)pyrimidin-2-amine

5-(6-chloro-2-morpholinopyrimidin-4-yl)pyrimidin-2-amine (10 mg, 0.034mmol, prepared as in example 1), potassium tert-butoxide (6 mg, 0.051mmol), pyridin-3-ol (5 mg, 0.051 mmol) and DMSO (0.5 mL) were allcombined together and heated at 110° C. for 2 days. The crude productwas purified directly by preparative reverse phase HPLC to give5-(2-morpholino-6-(pyridin-3-yloxy)pyrimidin-4-yl)pyrimidin-2-amine (5.1mg, 32%). LC/MS (m/z): 352.1 (MH⁺), R_(t) 1.83 minutes.

Example 9 Preparation of6-(2-aminopyrimidin-5-yl)-2-morpholino-N-(6-(piperazin-1-yl)pyridin-3-yl)pyrimidin-4-amine

To tert-butyl4-(5-(6-(2-aminopyrimidin-5-yl)-2-morpholinopyrimidin-4-ylamino)pyridin-2-yl)piperazine-1-carboxylate(prepared as described in Example 1 from5-(6-chloro-2-morpholinopyrimidin-4-yl)pyrimidin-2-amine andcommercially available tert-butyl4-(5-aminopyridin-2-yl)piperazine-1-carboxylate, 30 mg, 0.06 mmol) wereadded 5 mL of 4N HCl in dioxane. After stirring for one hour, thesolution was concentrated in vacuo. The residue was dissolved in a 3:1acetonitrile and water and lyophilized to afford the desired productLC/MS (m/z): 435.2 (MH⁺), R_(t) 1.52 minutes.

Example 10 Preparation of4-(trifluoromethyl)-5-(2,6-dimorpholinopyrimidin-4-yl)pyridin-2-amine

To a slurry of 2-morpholino-4,6-dichloropyrimidine (prepared as inMethod 22, 2.0 g, 8.54 mmol) in NMP (14 mL), triethylamine (1.43 mL,10.25 mmol) was added. The heterogeneous mixture was stirred for 15minutes, then treated with morpholine (0.75 mL, 8.54 mmol). Uponrefluxing at 85° C. under argon for 2 hours, the solution was cooled,then added to EtOAc (160 mL). The organic solution was washed with 25 mLof NaHCO_(3(sat.)) (2×), water (2×) and brine, dried over Na₂SO₄,filtered and concentrated. The crude material was dissolved in 200 mLEtOAc and filtered through a SiO₂ pad, further eluting with EtOAc,yielding 2.2 g (93%) of 2,4-dimorpholino-6-chloropyrimidine as anoff-white solid. LCMS (m/z): 285.0 (MH⁺), ¹H NMR (CDCl₃): δ 5.86 (s,1H), 3.71-3.76 (m, 12H), 3.52-3.56 (m, 4H).

4-(trifluoromethyl)-5-(2,6-dimorpholinopyrimidin-4-yl)pyridin-2-amine

Argon gas was bubbled through a heterogeneous mixture of2,4-dimorpholino-6-chloropyrimidine (4.1 g, 14.3 mmol) and4-(trifluoromethyl)-5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyridin-2-amine(16.5 g, 57.3 mmol) in 1,2-dimethoxyethane and 2M Na₂CO₃ (3:1) for 20minutes. 1,1′-Bis(diphenylphosphino)ferrocene palladium(II) chloride(292 mg, 0.36 mmol) was added and the high pressure glass vesselcontaining the mixture was sealed. The reaction mixture was then heatedat 90° C. for 15 hours, cooled and diluted with EtOAc (300 mL). Theorganic solution was washed with 300 mL of a mixture of water:Na₂CO_(3(sat.)):NH₄OH_((conc.))=5:4:1, then NH₄Cl_((sat)), and brine(2×), dried over Na₂SO₄, filtered and concentrated. The crude materialwas purified by SiO₂ chromatography (50-90% EtOAc/hexanes with 0.1% TEA)resulting in 5.62 g (95%) of4-(trifluoromethyl)-5-(2,6-dimorpholinopyrimidin-4-yl)pyridin-2-amine asan off-white solid. LCMS (m/z): 411.3 (MH⁺); ¹H NMR (CDCl₃): δ 8.27 (s,1H), 6.78 (s, 1H), 5.97 (s, 1H), 4.77 (bs, 2H), 3.59-3.80 (m, 12H),3.58-3.61 (m, 4H).

Example 11 Preparation ofN-(6-(1-isopropylpiperidin-4-yloxy)pyridin-3-yl)-6-(6-amino-4-(trifluoromethyl)pyridin-3-yl)-2-morpholinopyrimidin-4-amine

N-(6-(1-isopropylpiperidin-4-yloxy)pyridin-3-yl)-6-(6-amino-4-(trifluoromethyl)-pyridin-3-yl)-2-morpholinopyrimidin-4-aminewas synthesized according to the general procedure for the Buchwaldreaction in Example 2 by reacting6-(1-isopropylpiperidin-4-yloxy)pyridin-3-amine (prepared as in Method16) with5-(6-chloro-2-morpholinopyrimidin-4-yl)-4-(trifluoromethyl)pyridin-2-amine.LC/MS (m/z): 559.2 (MH⁺), R_(t) 1.92 minutes. ¹H NMR (DMSO): δ 10.27(1H, bs, NH); 8.41 (1H, bs); 8.17 (1H, s); 7.98 and 7.94 (1H, 2 bdoublets, J=9.0 Hz, 2 conformers); 6.97 (1H, s); 6.90 and 6.84 (1H, 2doublets, J=9.0 Hz, 2 conformers); 6.23 (1H, bs); 5.25 and 5.15 (1H, 2multiplets, 2 conformers); 3.66 (8H, bs); 3.44 (1H, m); 3.35 (2H, m);3.10 (2H, m); 2.22 (2H, m); 2.03 (2H, m); 1.27 (6H, overlapping doubletsbecause of conformers, app. triplet, J=5.7 Hz).

Example 12 Preparation ofN-(5-((diethylamino)methyl)thiazol-2-yl)-6-(6-amino-4-(trifluoromethyl)pyridin-3-yl)-2-morpholinopyrimidin-4-amine5-((diethylamino)methyl)thiazol-2-amine

2-Aminothiazole-5-carbaldehyde (1 eq) was added to a stirring solutionof diethylamine (4 eq) in anhydrous MeOH at 0° C. Sodiumcyanoborohydride (1.5 eq) was then added in portions at 0° C. Thereaction mixture was stirred at 70° C. for 10 hours. After this time,the solution was quenched with H₂O and extracted with EtOAc. Thecombined organic extracts were dried over Na₂SO₄ and concentrated toafford a viscous brown oil. LC/MS (m/z): 186.2 (MH⁺), R_(t) 0.33minutes.

The title compound was synthesized according to the general procedureshown in Example 2. LC/MS (m/z): 509.2 (MH⁺), R_(t) 1.98 minutes. ¹H NMR(DMSO): δ 11.0, (2H, bs, NH2), 8.17 (1H, s); 7.63 (1H, s); 7.08 (1H,bs), 6.40 (1H, s); 4.48 (2H, bd, J=4.2 Hz); 3.80 (4H, m); 168 (4H, m);3.03 (4H, bq, J=6.9 Hz); 1.30 (6H, t, J=6.9 Hz).

Example 13 Synthesis of6-(6-amino-4-(trifluoromethyl)pyridin-3-yl)-N-(4-(2-(diethylamino)ethyl)thiazol-2-yl)-2-morpholinopyrimidin-4-amine2-(2-aminothiazol-4-yl)-N,N-diethylacetamide

A mixture of 2-(2-aminothiazol-4-yl)acetic acid (1 eq), HOAT (1 eq), EDC(1.1 eq), TEA (1 eq) and HNEt₂ (1 eq) in DMA was stirred at roomtemperature for 6 hours. The reaction mixture was then quenched with H₂Oand concentrated. The residue was dissolved in a stirred 4:1 mixture ofEtOAc and NaHCO_(3(sat.)). The two phases were separated and the organicsolution was washed with brine, dried over Na₂SO₄ and concentrated todryness. The resulting solid was washed twice with Et₂O and dried toyield the desired product as a white solid. LC/MS (m/z): 214.0 (MH⁺),R_(t) 1.13 minutes.

4-(2-(diethylamino)ethyl)thiazol-2-amine

A suspension of 2-(2-aminothiazol-4-yl)-N,N-diethylacetamide (1 eq) inTHF was added dropwise to a vigorously stirred suspension of LAH (1 eq)in THF at 0° C. The mixture was stirred at room temperature for 2 hours.At this time, the resulting mixture was cooled to 0° C. and 1 part H₂O,followed by 1 part 10% NaOH and lastly 3 parts H₂O, were added dropwise.The mixture was stirred for 10 minutes, filtered, and the solid residuewashed with THF. The filtrate was collected and concentrated to dryness.The resulting crude material was washed with Et₂O twice and dried toafford a viscous brown oil. LC/MS (m/z): 200.1, (MH⁺), R_(t) 0.34minutes.

The title compound was synthesized according to the general procedureshown in Example 2. LC/MS (m/z): 523.1 (MH⁺), R_(t) 2.11 minutes. ¹H-NMR(DMSO): δ 8.15 (1H, s); 7.08 (1H, s); 6.96 (1H, s); 6.38 (1H, s); 3.78(4H, m); 3.65 (4H, m); 3.31 (2H, m); 3.13 (4H, q, J=7.2 Hz); 3.02 (2H,m); 1.20 (6H, t, J=7.2 Hz).

Example 14 Preparation ofN⁶-(2-methoxyethyl)-2-morpholino-4,5′-bipyrimidine-2′,6-diamine

An argon sparged mixture of6-chloro-2-morpholino-4,5′-bipyrimidin-2′-amine (10 mg, 0.03 mmol) and2-methoxyethanamine (0.018 mL, 0.20 mmol) in NMP (0.8 mL) contained in asealed pressure vessel was heated in a microwave at 155° C. for 1000seconds. The reaction mixture was filtered and purified by reverse phasepreparative HPLC to giveN⁶-(2-methoxyethyl)-2-morpholino-4,5′-bipyrimidine-2′,6-diamine as theTFA salt (4.0 mg, 30%). LC/MS (m/z): 332.2 (MH⁺), R_(t) 1.44 minutes.

Example 15 Preparation of2-morpholino-6-(2-phenylmorpholino)-4,5′-bipyrimidin-2′-amine

An argon sparged mixture of6-chloro-2-morpholino-4,5′-bipyrimidin-2′-amine (10 mg, 0.03 mmol),Cs₂CO₃ (27 mg, 0.09 mmol) and 2-phenylmorpholine (11 mg, 0.068 mmol) inNMP (0.5 mL) contained in a sealed pressure vessel, was heated in amicrowave at 170° C. for 600 seconds. The reaction mixture was filteredand purified by reverse phase preparative HPLC to give2-morpholino-6-(2-phenylmorpholino)-4,5′-bipyrimidin-2′-amine as the TFAsalt (7.2 mg, 45%). LC/MS (m/z): 420.1 (MH⁺), R_(t) 2.20 minutes.

Example 16 Preparation ofN⁶-tert-butyl-2-morpholino-4,5′-bipyrimidine-2′,6′-diamine

An argon sparged mixture of6-chloro-2-morpholino-4,5′-bipyrimidin-2′-amine (10 mg, 0.03 mmol) andtert-butylamine (12.5 mg, 0.17 mmol) in NMP (0.5 mL) contained in asealed pressure vessel, was heated in a microwave at 175° C. for 800seconds. An additional amount of tert-butylamine (50 mg, 0.68 mmol) wasadded to the reaction. The reaction was again heated in a microwave at175° C. for 800 seconds and again at 175° C. for 800 seconds untildisappearance of the starting material. The crude mixture was filtered.The crude product was purified by reverse phase preparative HPLC to giveN⁶-tert-butyl-2-morpholino-4,5′-bipyrimidine-2′,6-diamine as the TFAsalt (0.9 mg, 7%). LC/MS (m/z): 330.2 (MH⁺), R_(t) 1.96 minutes.

Example 17 Preparation of1-(2-(6-amino-4-(trifluoromethyl)pyridin-3-yl)-6-morpholino-pyrimidin-4-yl)piperidin-2-one5-(2-chloro-6-morpholinopyrimidin-4-ylamino)pentanoic acid

5-Aminopentanoic acid (140 mg, 1.19 mmol),4-(2,6-dichloropyrimidin-4-yl)morpholine (prepared as in Method 22; 234mg, 1.0 mmol) and DIEA (0.530 mL, 3.0 mmol) were dissolved inN,N-dimethylformamide (6 mL). The reaction solution was stirred at 40°C. for 40 hours. The reaction was diluted with EtOAc (100 mL) and washedwith 0.5 M HCl (40 mL), water (40 mL), brine (40 mL), dried with Na₂SO₄,filtered and evaporated to give a solid. The crude product waschromatographed on a silica gel column by eluting with 80% EtOAc inhexane to give 5-(2-chloro-6-morpholinopyrimidin-4-ylamino)pentanoicacid as a white solid (190 mg, 60%). LC/MS (m/z): 315.0 (MH⁺), R_(f)1.79 minutes.

1-(2-chloro-6-morpholinopyrimidin-4-yl)piperidin-2-one

To a solution of HATU (304 mg, 0.8 mmol), HOAT (82 mg, 0.6 mmol) andDIEA (0.209 mL, 1.2 mmol) in chloroform (20 mL) under argon, a solutionof 5-(2-chloro-6-morpholinopyrimidin-4-ylamino)pentanoic acid (190 mg,0.6 mmol) in chloroform (10 mL) was slowly added. The reaction solutionwas stirred at room temperature for 5 hours. After the reaction wascomplete, the solution was evaporated to dryness to give a white solidwhich was chromatographed on a silica gel column by eluting with 40%EtOAc/hexane to give1-(2-chloro-6-morpholinopyrimidin-4-yl)piperidin-2-one as a white solid(62 mg, 35%). LC/MS (m/z): 297.0 (MH⁺), R_(t) 2.74 minutes.

1-(2-(6-amino-4-(trifluoromethyl)pyridin-3-yl)-6-morpholinopyrimidin-4-yl)piperidin-2-one

To a suspension of1-(2-chloro-6-morpholinopyrimidin-4-yl)piperidin-2-one (16 mg, 0.05mmol),5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-4-(trifluoromethyl)pyridin-2-amine(prepared as in Method 4; 23 mg, 0.08 mmol) anddichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium(II)dichloromethane adduct (8 mg, 0.009 mmol) in dioxane (1.1 mL), 2 Maqueous sodium carbonate solution (0.4 mL, 0.8 mmol) was added underargon. The reaction mixture was heated in a microwave at 120° C. for1000 seconds. The crude product was partitioned between EtOAc (30 mL)and saturated sodium bicarbonate (10 mL). The organic layer wasseparated, dried over sodium sulfate, filtered and concentrated underreduced pressure. The crude product was purified by preparative reversephase HPLC to give1-(2-(6-amino-4-(trifluoromethyl)pyridin-3-yl)-6-morpholinopyrimidin-4-yl)piperidin-2-oneas a yellow powder (8.8 mg, 42%). LC/MS (m/z): 423.0 (MH⁺), R_(t) 2.25minutes.

Example 18 Preparation of1-(6-(6-amino-4-(trifluoromethyl)pyridin-3-yl)-2-morpholinopyrimidin-4-yl)-3-phenylimidazolidin-2-oneN¹-(6-chloro-2-morpholinopyrimidin-4-yl)-N²-phenylethane-1,2-diamine

To a solution of 4-(4,6-dichloropyrimidin-2-yl)morpholine (prepared asdescribed in Method 22; 932 mg, 4.0 mmol) and DMA (0.7 mL, 4.0 mmol) inACN (40 mL), neat N¹-phenyl-ethane-1,2-diamine (0.523 mL, 4.0 mmol) wasslowly added. The reaction mixture was stirred at 70-80° C. undernitrogen. After 20 hours, the reaction mixture was cooled down, and thesolvent was removed under reduced pressure. The crude product waspartitioned between EtOAc (120 mL) and 0.1 M NaHCO₃ (50 mL). The organiclayer was washed with additional 0.1 M NaHCO₃ (2×50 mL), brine (50 mL),dried, filtered and concentrated to giveN¹-(6-chloro-2-morpholinopyrimidin-4-yl)-N²-phenylethane-1,2-diamine, asan off-white solid (1.29 g, 96%). LC/MS (m/z): 334.0 (MH⁺), R_(t) 1.94minutes.

1-(6-chloro-2-morpholinopyrimidin-4-yl)-3-phenylimidazolidin-2-one

To a solution ofN¹-(6-chloro-2-morpholinopyrimidin-4-yl)-N²-phenylethane-1,2-diamine(100 mg, 0.3 mmol) in DCM (15 mL) at 0° C. under nitrogen, a solution ofphosgene in toluene (1.89 M, 0.32 mL, 0.6 mmol) was slowly added. After20 minutes, the reaction was allowed to warm to RT. After 18 hours, DIEA(0.42 mL, 2.4 mmol) was added and the reaction solution was heated to40-50° C. for 40 hours. The reaction mixture was evaporated underreduced pressure and the crude product was purified by silica gelchromatography eluting with 70% EtOAc/hexane to give1-(6-chloro-2-morpholinopyrimidin-4-yl)-3-phenylimidazolidin-2-one as awhite solid (94 mg, 87%). LC/MS (m/z): 360.1 (MH⁺), R_(t) 3.41 minutes.

1-(6-(6-amino-4-(trifluoromethyl)pyridin-3-yl)-2-morpholinopyrimidin-4-yl)-3-phenylimidazolidin-2-one

To a suspension of1-(6-chloro-2-morpholinopyrimidin-4-yl)-3-phenylimidazolidin-2-one (18mg, 0.05 mmol),5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-4-(trifluoromethyl)pyridin-2-amine(prepared as described in Method 4; 18 mg, 0.06 mmol) anddichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium (II)dichloromethane adduct (3.2 mg, 0.004 mmol) in DME (1.2 mL), 2 M aqueoussodium carbonate solution (0.4 mL, 0.8 mmol) was added under argon. Thereaction mixture was stirred at 95° C. for 5 hours. The crude productwas partitioned between EtOAc (30 mL) and saturated sodium bicarbonate(10 mL). The organic layer was separated, dried over sodium sulfate,filtered and concentrated under reduced pressure. The crude product waspurified by preparative HPLC to give1-(6-(6-amino-4-(trifluoromethyl)pyridin-3-yl)-2-morpholinopyrimidin-4-yl)-3-phenylimidazolidin-2-oneas a pale yellow powder (8.4 mg, 35% overall yield). LC/MS (m/z): 448.1(MH⁺), R_(t) 3.29 minutes.

Example 19 Preparation of1-(4-(6-(6-amino-4-(trifluoromethyl)pyridin-3-yl)-2-morpholinopyrimidin-4-yloxy)piperidin-1-yl)ethanoneStep 1: Alkoxylation of 2-morpholino-4,6-dichloropyrimidine

To a solution of N-Boc-4-hydroxy piperidine (2.58 g, 12.81 mmol) intetrahydrofuran at 0° C. under argon, was added sodium hydride (60%, 512mg, 12.81 mmol). After stirring for 20 minutes, a solution of2-morpholino-4,6-dichloropyrimidine (2.0 g, 8.54 mmol) intetrahydrofuran (20 mL) was added through a syringe. The solution wasstirred for 14 hours as the ice bath warmed to room temperature. At thistime, the reaction mixture was quenched with water (2 mL), and waspartitioned between EtOAc (350 mL) and Na₂CO₃(sat.) (75 mL). The organiclayer was separated, washed with brine (50 mL), dried over Na₂SO₄,filtered, concentrated and the residue was purified by SiO₂chromatography (15-20% EtOAc in hexanes) to yield tert-butyl4-(6-chloro-2-morpholinopyrimidin-4-yloxy)piperidine-1-carboxylate as awhite solid (2.64 g, 77%). LCMS (m/z): 399.1 (MH⁺). ¹H NMR (CDCl₃): δ6.00 (s, 1H), 5.18 (m, 1H), 3.74 (s, 8H), 3.64-3.74 (m, 2H), 3.28-3.38(m, 2H), 1.86-1.96 (m, 2H), 1.68-1.78 (m, 2H), 1.44 (s, 9H).

Step 2: Suzuki Reaction of2-morpholino-4-alkoxy-substituted-6-chloropyrimidine

A mixture of tent-butyl4-(6-chloro-2-morpholinopyrimidin-4-yloxy)piperidine-1-carboxylate (250mg, 0.63 mmol),5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-4-(trifluoromethyl)pyridine-2-amine(prepared as in method 4, 325 mg, 1.13 mmol) and Pd(dppf)Cl₂—CH₂Cl₂(25.6 mg, 0.031 mmol) in dimethoxyethane/2 M Na₂CO₃ (3:1, 8 mL) washeated under microwave irradiation for 15 minutes at 120° C. Thereaction mixture was partitioned between EtOAc (200 mL) andNa₂CO_(3(sat.)) (50 mL), the organic layer was separated, washed withbrine (50 mL), dried over Na₂SO₄, filtered, concentrated and purified bySiO₂ chromatography (50-75% EtOAc/hexanes) to yield the product as awhite solid (207 mg, 63%). LCMS (m/z): 525.2 (MH⁺).

Step 3: Hydrolysis of N-Boc Protecting Group

A mixture of tert-butyl4-(6-(6-amino-4-(trifluoromethyl)pyridin-3-yl)-2-morpholinopyrimidin-4-yloxy)piperidine-1-carboxylate(649 mg, 1.24 mmol) and 4 M HCl/dioxane (15 mL, 60 mmol) was allowed tostand at room temperature for 14 hours. Upon removal of volatiles invacuo, diethyl ether (50 mL) was added, the material was sonicated andconcentrated yielding the bis HCl salt of the desired product as an offwhite solid. LCMS (m/z): 425.1 (MH⁺).

Step 4: Acylation

To a solution of4-(trifluoromethyl)-5-(2-morpholino-6-(piperidin-4-yloxy)pyrimidin-4-yl)pyridin-2-aminein NMP, was added diisopropylethylamine (5 eq) and acetyl chloride (1.5eq). The reaction mixture was stirred at room temperature for 2 h andthen was purified directly by reverse-phase HPLC and lyophilizedyielding the TFA salt of the product. Alternatively, after reverse phaseHPLC the free base of the product could be isolated after extractioninto EtOAc upon basifying, followed by drying over Na₂SO₄ and removal ofvolatiles in vacuo. LCMS (m/z): 467.1 (MH⁺).

Example 20 Preparation of5-(6-((S)-piperidin-3-yloxy)-2-morpholinopyrimidin-4-yl)-4-(trifluoromethyl)pyridin-2-amine

(S)-tert-butyl3-(6-chloro-2-morpholinopyrimidin-4-yloxy)piperidine-1-carboxylate wasprepared as in Example 19, Step 1, for the alkoxylation of2-morpholino-4,6-dichloropyrimidine (87%). LCMS (m/z): 399.1 (MH⁺). TheBoc protected intermediate was prepared by Suzuki reaction as shown inStep 2 of Example 19 and was purified by SiO₂ chromatography (30-60%EtOAc/hexanes; 78%). LCMS (m/z): 526.0 (MH⁺). The title compound wasprepared by cleaving the N-Boc protecting group as shown in Step 3 ofExample 19. LCMS (m/z): 425.1 (MH⁺).

Example 21 Preparation of 5-(6-((R)-piperidin-3-yloxy)-2-morpholinopyrimidin-4-yl)-4-(trifluoromethyl)pyridin-2-amine

(R)-tert-butyl3-(6-chloro-2-morpholinopyrimidin-4-yloxy)piperidine-1-carboxylate wasprepared as in Example 19, Step 1, for the alkoxylation of2-morpholino-4,6-dichloropyrimidine (82%). LCMS (m/z): 399.1 (MH⁺). TheBoc protected intermediate was prepared by Suzuki reaction as shown inStep 2 of Example 19 and was purified by silica gel chromatography(30-60% EtOAc/hexanes, 54%). LCMS (m/z): 526.0 (MH⁺). The title compoundwas prepared by cleaving the N-Boc protecting group as shown in Step 3of Example 19. LCMS (m/z): 425.1 (MH⁺).

Example 22 Preparation of1-((R)-3-(6-(6-amino-4-(trifluoromethyl)pyridin-3-yl)-2-morpholinopyrimidin-4-yloxy)pyrrolidin-1-yl)ethanone

(R)-tert-butyl3-(6-chloro-2-morpholinopyrimidin-4-yloxy)pyrrolidine-1-carboxylate wasprepared as in Example 19, Step 1, for the alkoxylation of2-morpholino-4,6-dichloropyrimidine (41%). LCMS (m/z): 385.0 (MH⁺).

The Boc protected intermediate was prepared by Suzuki reaction as shownin Step 2 of Example 19 and was purified by reverse phase HPLC andisolated as free base after extraction into EtOAc upon basifying (71%).LCMS (m/z): 511.0 (MH⁺). Cleavage of the N-Boc protecting group wasperformed as shown in Step 3 of Example 19. LCMS (m/z): 411.0 (MH⁺). Thetitle compound was prepared as in Step 4 of Example 19. LCMS (m/z):453.1 (MH⁺), R_(t) 2.18.

Example 23 Preparation of1-((S)-3-(6-(6-amino-4-(trifluoromethyl)pyridin-3-yl)-2-morpholinopyrimidin-4-yloxy)pyrrolidin-1-yl)ethanone

(S)-tert-butyl3-(6-chloro-2-morpholinopyrimidin-4-yloxy)pyrrolidine-1-carboxylate wasprepared according to Example 19, Step 1, for the alkoxylation of2-morpholino-4,6-dichloropyrimidine (99%). LCMS (m/z): 385.0 (MH⁺). TheBoc protected intermediate was prepared by Suzuki reaction as shown inStep 2 of Example 19, purified by reverse phase HPLC and isolated asfree base after extraction into EtOAc upon basifying (72%). LCMS (m/z):511.0 (MH⁺). The N-Boc protecting group was cleaved as shown in Step 3of Example 19. LCMS (m/z): 411.0 (MH⁺). The title compound was preparedas in step 4 of Example 19. LCMS (m/z): 453.1 (MH⁺), R_(t) 2.18.

Example 24 Preparation of4-(trifluoromethyl)-5-(2-morpholino-6-(tetrahydro-2H-pyran-4-yloxy)pyrimidin-4-yl)pyridin-2-amine

4-(4-chloro-6-(tetrahydro-2H-pyran-4-yloxy)pyrimidin-2-yl)morpholine wasprepared according to Example 19, Step 1, for the alkoxylation of2-morpholino-4,6-dichloropyrimidine with 4-hydroxytetrahydropyran (80%).LCMS (m/z): 300.1 (MH⁺). The title compound was prepared by Suzukireaction as shown in Step 2 of Example 19. LC/MS (m/z): 426.1 (MH+),R_(t) 2.26 minutes.

Example 25 Preparation 5-(6-((R)-tetrahydrofuran-3-yloxy)-2-morpholinopyrimidin-4-yl)-4-(trifluoromethyl)pyridin-2-amine

(R)-4-(4-chloro-6-(tetrahydrofuran-3-yloxy)pyrimidin-2-yl)morpholine wasprepared according to Example 19, Step 1, for the alkoxylation of2-morpholino-4,6-dichloropyrimidine with (R)-3-hydroxytetrahydrofuran(81%). LCMS (m/z): 286.1 (MH⁺). The title compound was prepared bySuzuki reaction as shown in Step 2 of Example 19. LC/MS (m/z): 412.1(MH⁺).

Example 26 Preparation of5-(6-((S)-tetrahydrofuran-3-yloxy)-2-morpholinopyrimidin-4-yl)-4-(trifluoromethyl)pyridin-2-amine

(S)-4-(4-chloro-6-(tetrahydrofuran-3-yloxy)pyrimidin-2-yl)morpholine wasprepared according to Example 19, Step 1, for alkoxylation of2-morpholino-4,6-dichloropyrimidine with (S)-3-hydroxytetrahydrofuran(85%). LCMS (m/z): 286.1 (MH⁺). The title compound was prepared bySuzuki reaction as shown in Step 2 of Example 19. LC/MS (m/z): 412.1(MH⁺).

Example 27 Preparation of4-(trifluoromethyl)-5-(2-morpholino-6-(piperidin-4-yloxy)pyrimidin-4-yl)pyrimidin-2-amine

tert-butyl4-(6-(2-amino-4-(trifluoromethyl)pyrimidin-5-yl)-2-morpholinopyrimidin-4-yloxy)piperidine-1-carboxylatewas prepared by Suzuki reaction of tert-butyl4-(6-chloro-2-morpholinopyrimidin-4-yloxy)piperidine-1-carboxylate, asshown in Step 2 of Example 19, with5-(4,4,5,5-tetramethyl(1,3,2-dioxaborolan-2-yl))-4-(trifluoromethyl)pyrimidine-2-ylamine(prepared as in Method 5). The crude product was purified by silica gelchromatography (30-50% EtOAc/hexanes) (63%). LCMS (m/z): 526.0 (MH⁺).The title compound was prepared by cleaving the N-Boc protecting groupas shown in Step 3 of Example 19. LCMS (m/z): 426.0 (MH⁺).

Example 28 Preparation of5-(2-morpholino-6-(piperidin-4-yloxy)pyrimidin-4-yl)pyrimidine-2,4-diamine

tert-butyl4-(6-(2,4-diaminopyrimidin-5-yl)-2-morpholinopyrimidin-4-yloxy)piperidine-1-carboxylatewas prepared by Suzuki reaction tert-butyl4-(6-chloro-2-morpholinopyrimidin-4-yloxy)piperidine-1-carboxylate, asshown in Step 2 of Example 19, with5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidine-2,4-diamine(prepared as in Method 7). The crude product was purified by reversephase HPLC and isolated as the free base after extraction into EtOAcupon basifying (70%). LCMS (m/z): 473.1 (MH⁺). The title compound wasprepared by cleaving the N-Boc protecting group as shown in Step 3 ofExample 19. LCMS (m/z): 373.0 (MH⁺).

Example 29 Preparation of1-((R)-3-(6-(2,4-diaminopyrimidin-5-yl)-2-morpholinopyrimidin-4-yloxy)piperidin-1-yl)ethanone

(R)-tert-butyl3-(6-(2,4-diaminopyrimidin-5-yl)-2-morpholinopyrimidin-4-yloxy)piperidine-1-carboxylatewas prepared by Suzuki reaction of4-(6-chloro-2-morpholinopyrimidin-4-yloxy)piperidine-1-carboxylate, asshown in Step 2 of Example 19, with5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)pyrimidine-2,4-diamine.The crude product was purified by reverse phase HPLC and isolated as thefree base after extraction into EtOAc upon basifying (77%). LCMS (m/z):473.1 (MH⁺). The N-Boc protecting group was cleaved as shown in Step 3of Example 19. LCMS (m/z): 373.0 (MH⁺). The title compound wassynthesized as shown in step 4 of Example 19. LCMS (m/z): 460.1 (MH⁺),R_(t) 2.51.

Example 30 Preparation of2-amino-5-(2-morpholino-6-(N-acyl-piperidin-4-yloxy)pyrimidin-4-yl)pyrimidin-4(3H)-one

A mixture of tert-butyl4-(6-chloro-2-morpholinopyrimidin-4-yloxy)piperidine-1-carboxylate (500mg, 1.26 mmol), 4-methoxy-2-aminopyrimidyl boronate ester (prepared asin Method 8, 630 mg, 2.51 mmol) and Pd(dppf)Cl₂—CH₂Cl₂ (51 mg, 0.063mmol) in dimethoxyethane and 2 M Na₂CO₃ (3:1, 12 mL) was heated undermicrowave irradiation for 15 minutes at 120° C. The reaction mixture waspartitioned between EtOAc (200 mL) and Na₂CO_(3(sat.)) (50 mL), theorganic layer was separated and washed with brine (50 mL). The combinedaqueous layers were extracted further with EtOAc (2×100 mL), and thecombined organic layers were dried over Na₂SO₄, filtered andconcentrated. To this material was added 4 M HCl/dioxane (20 mL) toremove the Boc group. After standing for 12 hours, the volatiles wereremoved in vacuo, and the residue was partitioned between CH₂Cl₂ (200mL) and 1N NaOH (50 mL). Upon separating, the aqueous layer wasextracted further with CH₂Cl₂ (200 mL) and then CHCl₃ (2×150 mL). Thecombined organic layers were concentrated yielding1,6-dihydro-6-methoxy-5-(2-morpholino-6-(piperidin-4-yloxy)pyrimidin-4-yl)pyrimidin-2-amine(464 mg). The crude compound and morpholine (0.9 mL, 10.45 mmol) in NMP(10 mL) were heated under microwave irradiation for 15 minutes at 200°C. to convert the methoxy pyrimidine to the pyrimidone. Additionalmorpholine (0.9 mL, 10.45 mmol) was added and the solution was heatedunder microwave irradiation for 15 minutes and than 10 minutes at 200°C. Upon cooling the material was directly purified by reverse-phaseHPLC. After lyophilization, the bis TFA salt of the2-amino-5-(2-morpholino-6-(piperidin-4-yloxy)pyrimidin-4-yl)pyrimidin-4(3H)-onewas isolated as an off-white solid (325 mg, 45%). LCMS (m/z): 374.1(MH⁺). The title compound was prepared by acylation of the secondaryamino group as shown in Example 19, step 4. LCMS (m/z): 416.0 (MH⁺),R_(t) 1.67.

Example 31 Preparation of2-amino-5-(2-morpholino-6-(N-methoxycarbonyl-piperidin-4-yloxy)pyrimidin-4-yl)pyrimidin-4(3H)-one

The title compound was prepared as in Example 30, except utilizingmethylchloroformate instead of acetyl chloride in the last step. LCMS(m/z): 432.0 (MH⁺), R_(t) 2.05.

Example 32 Preparation of6-[6-amino-4-(trifluoromethyl)pyridin-3-yl]-N-[4-(1-isopropylpiperidin-4-yloxy)phenyl]-2-morpholinopyrimidin-4-amine

In a glass pressure vessel, Pd(OAc)₂ (5.0 mg, 0.022 mmol), BINAP (17.0mg, 0.028 mmol), cesium carbonate (72.0 mg, 0.22 mmol) and THF (2.0 mL)were mixed and stirred at room temperature for 1-3 minutes. To theresulting mixture was added5-(6-chloro-2-morpholin-4-yl-pyrimidin-4-yl)-pyridin-2-ylamine (40.0 mg,0.11 mmol) followed by 4-(1-isopropylpiperidin-4-yloxy) aniline (37.0mg, 0.16 mmol). The glass pressure vessel was sealed, stirred, andheated in microwave under irradiation at 110° C. for 10 minutes. Thereaction mixture was filtered and concentrated under reduced pressure.The product was purified by preparative reverse phase HPLC to give6-[6-amino-4-(trifluoromethyl)pyridin-3-yl]-N-[4-(1-isopropylpiperidin-4-yloxy)phenyl]-2-morpholinopyrimidin-4-amine(3.0 mg, 5%). LC/MS (m/z): 558.3 (MH⁺), R_(t) 1.90 minutes.

Example 33 Preparation of6-(6-amino-4-(trifluoromethyl)pyridin-3-yl)-N-(4-(1-isopropylpiperidin-4-yloxy)-3-methoxyphenyl)-2-morpholinopyrimidin-4-amine

In a glass pressure vessel, Pd(OAc)₂ (5.0 mg, 0.02 mmol), BINAP (17.0mg, 0.028 mmol), cesium carbonate (72.0 mg, 0.22 mmol) and THF (2.0 mL)were mixed and stirred at room temperature for 1-3 minutes. To theresulting mixture was added5-(6-chloro-2-morpholin-4-yl-pyrimidin-4-yl)-pyridin-2-ylamine (40.0 mg,0.11 mmol) followed by 4-(1-isopropylpiperidin-4-yloxy)-3-methoxyaniline(46.6 mg, 0.16 mmol). The glass pressure vessel was sealed, stirred, andheated in microwave under irradiation at 120° C. for 10 minutes. Thereaction mixture was filtered and concentrated under reduced pressure.The residue was purified by preparative reverse phase HPLC to give6-(6-amino-4-(trifluoromethyl)pyridin-3-yl)-N-(4-(1-isopropylpiperidin-4-yloxy)-3-methoxyphenyl)-2-morpholinopyrimidin-4-amine(6.6 mg, 10%). LC/MS (m/z): 588.3 (MH⁺), R_(t) 1.92 minutes.

Example 34 Synthesis ofN-(6-(6-amino-4-(trifluoromethyl)pyridin-3-yl)-2-morpholinopyrimidin-4-yl)-4-phenylthiazol-2-amine

A solution ofN-(6-chloro-2-morpholinopyrimidin-4-yl)-4-phenylthiazol-2-amine (15 mg,0.040 mmol),5-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-4-(trifluoromethyl)pyridin-2-amine(23 mg, 0.080 mmol) and 1,1′-bis(diphenylphosphino)ferrocene palladium(II) chloride (6.6 mg, 0.0080 mmol) in 0.5 mL of 1,4-dioxane and 0.05 mLof 2 M aq. sodium carbonate was heated in the microwave at 120° C. for600 seconds. The crude product was purified by reverse phase prep HPLCto giveN-(6-(6-amino-4-(trifluoromethyl)pyridin-3-yl)-2-morpholinopyrimidin-4-yl)-4-phenylthiazol-2-amine.LC/MS (m/z): 500 (MH⁺), R_(t) 2.46 minutes.

Examples 35 Preparation ofN-6-(6-amino-4-(trifluoromethyl)pyridin-3-yl)-5-methyl-2-morpholinopyrimidin-4-yl)-4-phenylthiazol-2-amine

N-(6-(6-amino-4-(trifluoromethyl)pyridin-3-yl)-5-methyl-2-morpholinopyrimidin-4-yl)-4-phenylthiazol-2-aminewas prepared according to Example 35. LC/MS (m/z): 514 (MH⁺), R_(t) 2.62minutes.

TABLE 1 hplc PI3 Cellular LC/MS 10 min Kinase PSer473 Prolifer- Com-(M + H, (L = Alpha, AKT, ation, pound Structure Rt min) 45 min) IC₅₀EC₅₀ EC₅₀ 1

401.4, 2.00 ++++ ++++ ++++ 2

380.1, 1.82 9.67 L ++++ N/D N/D 3

428.2, 2.09 11.50 L ++++ ++++ ++++ 4

418.0 1.99 ++++ ++++ ++++ 5

425.0 11.16 L ++++ +++ ++++ 6

372.2 ++++ N/D +++ 7

431.2, 2.03 ++++ ++++ ++++ 8

352.1, 1.83 ++++ ++++ ++++ 9

435.2, 1.52 ++++ ++++ ++++ 10

411.3, 1.86 ++++ ++++ ++++ 11

559.2, 1.92 ++++ ++++ ++++ 12

509.0, 1.72 ++++ ++++ +++ 13

523.1, 2.02 2.11 ++++ ++++ ++++ 14

332.2 7.50 L ++++ N/D ++++ 15

420.1 13.14 L ++++ ++++ ++++ 16

330.2 10.83 L ++++ N/D ++++ 17

423.1 2.57 ++++ N/D ++++ 18

486 3.23 ++++ N/D ++++ 19

467.1, 2.36 ++++ N/D ++++ 20

525.0, 3.42 ++++ N/D ++++ 21

525.0, 3.42 ++++ N/D N/D 22

453.1, 2.18 ++++ N/D +++ 23

453.1, 2.18 ++++ N/D N/D 24

426.1, 2.26 2.54 ++++ ++++ ++++ 25

412.1, 2.47 ++++ ++++ +++ 26

412.1, 2.19 2.47, (12.34) ++++ N/D +++ 27

526.0, 4.30 ++++ ++++ ++++ 28

473.1, 3.02 ++++ ++++ ++++ 29

415.1, 2.06 ++++ ++++ ++++ 30

416.0, 1.67 ++++ ++++ ++++ 31

432.0, 2.05 ++++ ++++ ++++ 32

460.1, 2.51 ++++ ++++ ++++ 33

419.0, 2.17 ++++ ++++ ++++ 34

508.0, 2.17 2.96, (14.82) ++++ ++++ ++++ 35

450.2, 1.61 ++++ ++++ ++++ 36

438.1, 1.61 ++++ ++++ ++++ 37

464.4, 1.53 ++++ ++++ ++++ 38

402.2, 1.88 min ++++ ++++ ++++ 39

361.0, 1.44 1.73, (8.13) ++++ ++++ ++++ 40

510.1, 1.98 2.18 ++++ ++++ ++++ 41

560.2, 1.93 1.98 ++++ ++++ ++++ 42

478.4, 1.59 ++++ ++++ ++++ 43

4.19 9.23 L ++++ ++++ ++++ 44

351.1 8.23 L ++++ ++++ ++++ 45

375.0, 2.11 2.41 ++++ ++++ ++++ 46

401.1, 1.70 1.70 ++++ ++++ ++++ 47

486.1, 2.11 2.76, (14.49) ++++ ++++ ++++ 48

478.0, 1.79 2.23, (11.48) ++++ ++++ ++++ 49

444.1, 1.70 2.06, (10.23) ++++ ++++ ++++ 50

375.1, 1.80 1.82, (8.95) ++++ ++++ ++++ 51

449.1, 1.51 6.56 L ++++ ++++ ++++ 52

433.0, 2.30 ++++ ++++ ++++ 53

380.1, 1.65 ++++ ++++ ++++ 54

498.9, 2.55 ++++ ++++ ++++ 55

524.1, 2.10 2.37 ++++ ++++ ++++ 56

361.0, 1.45 1.64, (8.18) ++++ ++++ ++++ 57

366.1, 1.85 1.95 ++++ ++++ ++++ 58

452.0, 1.65 ++++ ++++ ++++ 59

457.2, 1.72 1.71 ++++ ++++ ++++ 60

431.2, 1.95 10.48 L ++++ ++++ ++++ 61

247.4, 2.85 ++++ ++++ ++++ 62

532.0, 1.85 ++++ ++++ ++++ 63

431.2, 2.43 ++++ ++++ ++++ 64

444.4, 1.66 ++++ ++++ ++++ 65

392.3, 2.55 ++++ ++++ ++++ 66

427.1, 3.21 ++++ ++++ +++ 67

408.1, 1.98 2.16 ++++ ++++ ++++ 68

507.2, 1.79 1.78 ++++ ++++ ++++ 69

496.9, 2.40 3.39, (16.57) ++++ ++++ ++++ 70

484.0, 3.36 ++++ ++++ ++++ 71

396.3, 2.32 ++++ ++++ ++++ 72

443 .2, 2.45 ++++ ++++ +++ 73

396.1, 1.58 1.89, (9.53) ++++ ++++ +++ 74

504.0, 3.19 ++++ ++++ ++++ 75

431.2, 2.38 ++++ ++++ +++ 76

359.1, 1.42 ++++ ++++ +++ 77

360.0, 1.47 1.79, (8.61) ++++ ++++ ++++ 78

496.9, 2.42 ++++ ++++ ++++ 79

544.2, 1.97 2.04 ++++ ++++ +++ 80

410.1, 1.91 10.36 L ++++ ++++ ++++ 81

431.0, 2.45 ++++ ++++ ++++ 82

400.0, 1.74 1.76 ++++ ++++ ++++ 83

529.2, 2.98 ++++ ++++ +++ 84

374.1, 2.13 ++++ ++++ ++++ 85

412.0, 2.03 ++++ ++++ ++++ 86

545.6, 1.78 ++++ ++++ ++++ 87

368.1, 2.05 2.26 ++++ ++++ +++ 88

502.1, 1.89 1.95 ++++ ++++ +++ 89

451.1, 2.30 ++++ ++++ ++++ 90

496 2.29 ++++ ++++ ++++ 91

381.4, 1.95 ++++ ++++ ++++ 92

437.1, 2.33 2.8 ++++ ++++ ++++ 93

545.6, 1.78 ++++ ++++ ++++ 94

443.1 2.07 ++++ ++++ ++++ 95

480.4, 2.13 2.85, (14.41) ++++ ++++ ++++ 96

415.3, 1.90 ++++ ++++ ++++ 97

485.1, 3.04 ++++ ++++ ++++ 98

429.2 8.99 L ++++ ++++ ++++ 99

415.1, 1.97 ++++ ++++ ++++ 100

344.1 7.58 L ++++ ++++ ++++ 101

360.1, 2.05 ++++ ++++ +++ 102

394.2, 1.85 1.92 ++++ ++++ ++++ 103

415.3, 1.68 ++++ ++++ ++++ 104

394.4, 1.62 ++++ ++++ +++ 105

428.9, 1.72 2.25, (10.78) ++++ ++++ ++++ 106

496.0, 3.28 ++++ ++++ ++++ 107

443.4, 270 ++++ ++++ +++ 108

360.1, 2.05 ++++ ++++ ++++ 109

496.0, 2.07 2.39 ++++ ++++ +++ 110

432.1 1.97 ++++ ++++ ++++ 111

368.0; 2.15 2.48 ++++ ++++ +++ 112

427.1 2.08 ++++ ++++ +++ 113

501.1, 2.34 14.12 L ++++ ++++ +++ 114

3.77.0, 1.54 ++++ ++++ +++ 115

512.0, 3.96 ++++ ++++ +++ 116

531.5, 1.77 ++++ ++++ +++ 117

483.1, 2.37 2.82, (14.09) ++++ ++++ +++ 118

383.0, 2.76 2.53 ++++ ++++ ++++ 119

468.0, 2.70 ++++ ++++ ++++ 120

451.0, 2.28 ++++ ++++ +++ 121

487.9 3.45 ++++ ++++ ++++ 122

477.1, 2.60 ++++ ++++ ++++ 123

368.1, 2.09 2.34 ++++ ++++ +++ 124

530.0, 3.53 ++++ ++++ ++++ 125

428.0, 2.38 ++++ ++++ ++++ 126

466.1, 2.25 2.62 ++++ ++++ +++ 127

451.1, 2.25 ++++ ++++ +++ 128

496.1 2.26 ++++ ++++ +++ 129

530.1, 1.93 1.99 ++++ ++++ +++ 130

364.1, 1.69 1.76 ++++ ++++ ++++ 131

459.1, 2.82 ++++ ++++ ++++ 132

259.2, 1.27 1.34, (6.23) ++++ ++++ +++ 133

471.2, 1.79 1.88 ++++ ++++ ++++ 134

443.2, 2.37 ++++ ++++ ++++ 135

390.1, 1.85 9.52 L ++++ ++++ ++++ 136

453.0, 2.29 2.76 ++++ ++++ ++++ 137

443.2, 2.38 ++++ ++++ +++ 138

409.0, 2.95 ++++ ++++ +++ 139

427.1 2.03 ++++ ++++ +++ 140

498.5, 2.36 ++++ ++++ +++ 141

427 2.38 ++++ ++++ +++ 142

418.1, 1.78 8.81 L ++++ ++++ ++++ 143

480.9, 2.46 3.50, (17.16) ++++ ++++ ++++ 144

448.9, 2.76 ++++ ++++ +++ 145

496.0, 2.35 2.75 ++++ ++++ +++ 146

507.2, 3.12 ++++ ++++ ++++ 147

416.0, 1.98 ++++ ++++ ++++ 148

380.1 1.78 ++++ ++++ ++ 149

478.9, 1.75 ++++ ++++ +++ 150

517.5, 1.78 ++++ ++++ ++++ 151

449.0, 2.42 ++++ ++++ +++ 152

375.0 2.22 ++++ ++++ +++ 153

486.4, 2.12 ++++ ++++ ++++ 154

445.3, 2.02 ++++ ++++ ++++ 155

384.0, 2.04 2.28 ++++ ++++ +++ 156

430.2, 2.05 11.18 L ++++ ++++ ++++ 157

478.1, 2.40 14.67 L ++++ ++++ ++ 158

486.9, 2.48 ++++ ++++ ++++ 159

495.0, 2.57 3.13 ++++ ++++ +++ 160

358.1 8.00 L ++++ ++++ +++ 161

431.4, 1.96 ++++ ++++ ++++ 162

368.2 1.84 ++++ ++++ ++++ 163

416.1 2.23 ++++ ++++ +++ 164

475.9, 2.69 ++++ ++++ ++++ 165

445.9, 1.95 2.57, (12.84) ++++ ++++ ++++ 166

392.1 1.62 ++++ ++++ +++ 167

459.2, 2.71 ++++ ++++ +++ 168

389.1 2.38 ++++ ++++ ++++ 169

418.3, 1.70 2.16, (10.66) ++++ ++++ ++++ 170

462.9, 2.41 ++++ ++++ ++++ 171

498.1 1.92 ++++ ++++ ++++ 172

391.2 2.62 ++++ ++++ +++ 173

495.0 2.32 ++++ ++++ +++ 174

391.1 2.14 ++++ ++++ +++ 175

434.3, 1.95 ++++ ++++ ++++ 176

560.0, 4.28 ++++ ++++ +++ 177

445.3, 1.79 ++++ ++++ ++++ 178

462.0, 1.98 2.19 ++++ ++++ +++ 179

374.0, 2.16 2.48 ++++ ++++ +++ 180

434.1 2.4 ++++ ++++ +++ 181

513.1, 1.76 1.72 ++++ ++++ ++++ 182

497.2, 1.90 9.89 L ++++ ++++ ++++ 183

400.4, 2.03 ++++ ++++ ++++ 184

350.1 7.65 L ++++ ++++ ++++ 185

429.4, 1.67 ++++ ++++ ++++ 186

485.1, 2.91 ++++ ++++ +++ 187

428.1 2.17 ++++ ++++ +++ 188

545.2, 1.84 1.89 ++++ ++++ +++ 189

379.4, 1.42 ++++ ++++ +++ 190

428.1 1.85 ++++ ++++ +++ 191

375.1, 1.75 1.74 ++++ ++++ +++ 192

469.4, 2.44 ++++ ++++ ++++ 193

468.4, 2.26 ++++ ++++ ++++ 194

524.5, 2.44 3.44 ++++ ++++ +++ 195

434.3, 2.06 ++++ ++++ ++++ 196

368.1, 1.69 1.63 ++++ ++++ ++++ 197

392.1 1.68 ++++ ++++ +++ 198

437.2, 1.60 1.45 ++++ ++++ N/D 199

448.4, 2.24 ++++ ++++ +++ 200

430.1, 1.84 9.55 L ++++ ++++ ++++ 201

414.1, 1.85 9.53 L ++++ ++++ ++++ 202

418.3 2.16 ++++ ++++ ++++ 203

491.1, 1.85 1.83 ++++ ++++ +++ 204

476.1, 2.42 2.86 ++++ ++++ ++ 205

316.2, 1.45 ++++ ++++ +++ 206

431.0, 1.91 2.16 ++++ ++++ ++++ 207

564.1 3.08 ++++ +++ ++++ 208

445.0, 1.50 1.78 (8.91) ++++ +++ ++ 209

351.0, 2.12 2.88, (14.36) ++++ +++ +++ 210

459.4, 2.80 ++++ +++ +++ 211

430.2, 2.02 2.14 ++++ +++ ++++ 212

455.5, 1.53 ++++ +++ ++ 213

428.2, 1.74 8.55 L ++++ +++ ++++ 214

444.0, 2.06 ++++ +++ ++ 215

524.5, 2.44 3.36 ++++ +++ +++ 216

487.9, 3.60 ++++ N/D +++ 217

469.1, 2.01 2.13 ++++ N/D +++ 218

537.1, 2.27 2.53 ++++ N/D +++ 219

452.0, 1.85 1.89 ++++ N/D +++ 220

494.1, 1.67 1.59 +++ N/D ++ 221

425.0, 1.66 1.98 ++++ N/D +++ 222

479.1, 1.98 2.20 +++ N/D N/D 223

423.2, 1.83 1.99 ++++ N/D +++ 224

370.3, 1.25 1.39 +++ N/D N/D 225

422.2, 1.84 1.86 ++++ N/D N/D 226

448.3, 1.93 1.94 ++++ N/D N/D 227

389.2, 1.93 1.93 ++++ N/D +++ 228

353.1, 2.25 2.55 ++++ N/D N/D 229

302.1, 1.68 1.74 ++++ N/D N/D 230

379.1, 1.73 1.74 ++++ N/D +++ 231

379.1, 1.75 1.78 ++++ N/D +++ 232

316.1, 1.84 2.24 ++++ N/D ++ 233

371.2, 1.49 1.39 ++++ N/D +++ 234

370.0, 2.12 2.37 ++++ N/D ++ 235

385.2, 1.50 1.40 ++++ N/D +++ 236

303.1, 1.65 1.70 ++++ N/D N/D 237

284.1, 2.12 2.56 ++++ N/D +++ 238

495.0 2.3 ++++ N/D +++ 239

443.1 2.63 ++++ N/D +++ 240

495, 2.20 3.29 ++++ N/D N/D 241

452.0 2.57 ++++ N/D +++ 242

461.1 1.85 ++++ N/D +++ 243

486, 2.01 2.26 ++++ N/D N/D 244

413, 2.13 2.41 ++++ N/D +++ 245

350, 1.72 1.66 ++++ N/D +++ 246

335, 1.66 1.57 ++++ N/D +++ 247

338, 1.92 2.02 ++++ N/D +++ 248

324, 1.79 1.82 ++++ N/D +++ 249

427, 2.15 2.40 ++++ N/D +++ 250

391, 2.07 2.30 ++++ N/D +++ 251

377, 1.98 2.14 ++++ N/D +++ 252

376, 2.31 2.66 ++++ N/D +++ 253

391, 2.13 2.56 ++++ N/D +++ 254

377, 1.76 1.81 ++++ N/D ++ 255

376, 2.14 2.39 ++++ N/D ++ 256

316, 1.44 1.32 ++++ N/D N/D 257

315, 1.46 1.30 +++ N/D N/D 258

274, 1.40 1.22 ++++ N/D ++ 259

273, 1.40 1.23 +++ N/D ++ 260

444.1, 2.02 2.24 ++++ N/D +++ 261

507.2, 1.92 1.98 ++++ N/D +++ 262

559.2, 2.07 2.25 ++++ N/D +++ 263

500.2, 1.66 2.03 ++++ N/D +++ 264

486.1, 1.61 1.94 ++++ N/D +++ 265

511.1, 2.09 2.44 ++++ N/D +++ 266

481.1, 2.23 2.59 ++++ N/D +++ 267

518.2, 2.18 ++++ N/D +++ 268

504.1, 2.13 ++++ N/D +++ 269

505.2, 1.76 ++++ N/D +++ 270

437.1, 1.56 1.44 ++++ N/D ++ 271

440.1 1.59 1.45 ++++ N/D ++ 272

529.1 1.64 1.72 ++++ N/D N/D 273

447.2, 1.61 1.54 ++++ N/D +++ 274

352.2, 1.81 1.77 ++++ N/D ++++ 275

365.2, 1.88 1.90 ++++ N/D ++++ 276

463.3, 1.72 ++++ N/D +++ 277

449.2, 2.00 2.11 ++++ N/D +++ 278

354.2, 2.32 2.12 ++++ N/D +++ 279

352.1, 1.81 1.48 ++++ N/D +++ 280

386.1, 1.83 1.91 ++++ N/D +++ 281

485.1, 2.17 ++++ N/D ++ 282

486.0, 1.69 ++++ N/D ++ 283

442.0, 2.02 ++++ N/D +++ 284

443.1, 2.22 ++++ N/D +++ 285

462.1, 1.95 ++++ N/D +++ 286

513.1, 2.46 +++ N/D N/D 287

430.1, 2.98 ++++ N/D +++ 288

434.4, 1.97 +++ N/D N/D 289

399.4, 1.50 ++++ N/D +++ 290

372.3, 1.74 ++++ N/D +++ 291

393.4, 1.32 ++++ N/D ++ 292

357.2, 1.78 +++ N/D N/D 293

371.4, 1.68 +++ N/D N/D 294

367.3, 1.65 ++++ N/D +++ 295

367.3, 2.17 ++++ N/D +++ 296

356.3, 1.22 +++ N/D N/D 297

378.4, 1.72 ++++ N/D N/D 298

383.4, 2.69 ++++ N/D N/D 299

434.5, 1.41 ++++ N/D +++ 300

448.4, 1.44 ++++ N/D +++ 301

274.2, 0.46 ++++ N/D ++ 302

407.2, 2.04 3.73 +++ N/D ++ 303

407.2, 2.02 3.77 ++++ N/D +++ 304

407.1, 2.10 2.25 ++++ N/D ++ 305

367.0; 2.07 2.28 ++++ N/D +++ 306

380.1; 2.07 2.29 ++++ N/D +++ 307

375.0; 2.09 2.39 ++++ N/D +++ 308

380.1; 2.07 2.32 ++++ N/D +++ 309

326.1, 1.79 2.99 ++++ N/D N/D 310

325.0, 1.51 1.88 ++++ N/D N/D 311

460.1, 1.96 2.09 ++++ N/D ++++ 312

445.1, 2.30 2.7 ++++ N/D N/D 313

429.1 2.32 ++++ N/D ++ 314

516.1 1.78 ++++ N/D ++ 315

579.1 2.09 ++++ N/D +++ 316

566 2.64 ++++ N/D +++ 317

400.1, 2.02 2.27 ++++ N/D +++ 318

525.1 2.15 ++++ N/D +++ 319

465.1, 2.28 2.5 ++++ N/D N/D 320

454.1, 1.74 1.74 ++++ N/D +++ 321

426.1 2.08 ++++ N/D +++ 322

425.1, 1.92 1.97 ++++ N/D ++ 323

425.0, 1.78 1.83 ++++ N/D ++ 324

423.0, 1.82 1.79 ++++ N/D +++ 325

524.1, 1.88 1.96 ++++ N/D ++ 326

482.1, 1.88 1.93 ++++ N/D ++ 327

439.2, 2.15 2.38 ++++ N/D +++ 328

392.0, 2.08 2.26 ++++ N/D ++ 329

538.2, 1.90 1.98 ++++ N/D ++ 330

496.2, 2.04 2.2 ++++ N/D ++ 331

459.1, 1.83 1.89 ++++ N/D ++ 332

413.1, 2.04 2.21 ++++ N/D ++++ 333

455.1, 1.77 1.79 ++++ N/D ++ 334

555.1, 2.76 3.36 ++++ N/D +++ 335

505.1 2.94 ++++ N/D +++ 336

521.1, 2.66 3.18 ++++ N/D +++ 337

521.1 3.1 ++++ N/D ++ 338

488.1, 1.73 1.67 ++++ N/D +++ 339

387.1, 1.55 1.44 +++ N/D N/D 340

420.1, 1.57 1.44 ++++ N/D N/D 341

444.1 2.84 ++++ N/D +++ 342

453.1 2.51 ++++ N/D +++ 343

488.1 3.02 ++++ N/D ++ 344

487.2 2.86 ++++ N/D +++ 345

389.1, 2.06 2.28 ++++ N/D +++ 346

389.1, 1.92 1.94 ++++ N/D N/D 347

389.1 1.83 ++++ N/D +++ 348

393.1 1.57 ++++ N/D ++ 349

384.1 2.13 ++++ N/D ++ 350

418.1 2.77 ++++ N/D ++ 351

368.2 1.77 ++++ N/D ++ 352

392.1, 1.89 1.94 ++++ N/D +++ 353

406.1, 1.78 1.77 +++ N/D N/D 354

432.2, 2.05 2.25 ++++ N/D N/D 355

415.0, 1.73 1.61 ++++ N/D N/D 356

432.0 2.0 ++++ N/D +++ 357

416.0, 2.05 2.21 ++++ N/D ++ 358

481.1, 2.64 3.27 ++++ N/D +++ 359

391.1, 2.06 2.28 ++++ N/D ++++ 360

406.1, 1.71 1.71 ++++ N/D +++ 361

442.1 1.89 ++++ N/D +++ 362

428.1 1.77 ++++ N/D +++ 363

406.1 1.77 ++++ N/D +++ 364

375.1, 1.93 2.04 ++++ N/D ++ 365

414.1, 1.94 10.78 L +++ N/D N/D 366

425.1, 2.14 12.06 L +++ N/D N/D 367

416.1 9.23 L ++++ N/D +++ 368

371.2, 1.69 7.86 L ++++ N/D +++ 369

292.1, 2.07 11.31 L ++++ N/D ++ 370

301.2, 1.57 6.77 L +++ N/D N/D 371

419.2 12.26 L ++++ N/D +++ 372

369.2, 2.15 11.91 L +++ N/D N/D 373

355.2, 2.07 11.27 L +++ N/D N/D 374

357.2, 1.62 7.19 L +++ N/D N/D 375

389.2, 2.13 12.07 L ++++ N/D N/D 376

356.2, 1.40 5.75 L +++ N/D N/D 377

401.1 10.23 L ++++ N/D +++ 378

350.2, 1.66 7.63 L ++++ N/D +++ 379

417.1, 2.28 13.32 L ++++ N/D N/D 380

4.68.1, 2.16 11.42 L ++++ N/D +++ 381

420.1, 1.81 9.41 L ++++ N/D N/D 382

389.2, 2.28 13.47 L: ++++ N/D N/D 383

468.2, 2.13 11.64 L ++++ N/D +++ 384

420.1, 1.68 8.71 L ++++ N/D +++ 385

418.1, 1.98 11.04 L ++++ N/D N/D 386

407.1, 1.95 10.57 L ++++ N/D +++ 387

391.1, 2.25 13.62 L ++++ N/D +++ 388

409.1, 1.87 9.91 L ++++ N/D N/D 389

407.1, 2.08 11.36 L ++++ N/D N/D 390

419.1 10.41 L ++++ N/D +++ 391

410.1, 2.20 12.63 L ++++ N/D N/D 392

357.1 5.96 L ++++ N/D +++ 393

418.1 13.00 L ++++ N/D +++ 394

404.2 10.64 L ++++ N/D +++ 395

462.2, 2.38 14.83 L ++++ N/D N/D 396

448.2 14.53 L ++++ N/D +++ 397

462.2, 2.40 14.82 L ++++ N/D N/D 398

448.2, 2.38 14.52 L ++++ N/D N/D 399

328.2 9.63 L ++++ N/D +++ 400

302.2 7.77 L ++++ N/D +++ 401

288.2 6.92 L ++++ N/D ++ 402

314.2 8.39 L ++++ N/D +++ 403

390.1 13.44 L ++++ N/D +++ 404

370.2 13.71 L ++++ N/D +++ 405

356.2 12.73 L ++++ N/D +++ 406

370.2 14.24 L ++++ N/D ++ 407

418.2 14.81 L ++++ N/D +++ 408

469.1 12.14 L ++++ N/D +++ 409

469.1 12.17 L ++++ N/D ++++ 410

469.1 12.17 L ++++ N/D +++ 411

390.1 13.47 L ++++ N/D +++ 412

421.1 8.70 L ++++ N/D ++++ 413

421.1 9.60 L ++++ N/D +++ 414

480.0, 1.98 2.19 ++++ N/D N/D 415

283.2, 1.95 ++++ N/D ++ 416

500.0, 1.83 2.36 ++++ N/D N/D 417

378.0, 3.02 2.79 ++++ N/D ++ 418

284.2, 1.94 2.2 ++++ N/D ++ 419

356.2, 1.77 1.86 +++ N/D N/D 420

373.2, 1.37 1.23 +++ N/D N/D 421

386.2, 1.57 1.73 +++ N/D N/D 422

330.2, 1.47 1.58 +++ N/D N/D 423

387.2, 1.47 1.32 +++ N/D N/D 424

360.2, 1.57 1.47 +++ N/D N/D 425

356.2, 1.79 1.83 +++ N/D N/D 426

386.2, 1.61 1.56 +++ N/D N/D 427

385.2, 1.48 1.4 +++ N/D ++ 428

385.2, 0.5 1.34 +++ N/D ++ 429

372.3, 1.39 1.69 +++ N/D N/D 430

303.1, 1.66 1.66 +++ N/D N/D 431

317.2, 1.59 2.02 ++++ N/D ++ 432

476.1, 2.16 2.46 ++++ N/D +++ 433

378.1, 1.31 1.13 +++ N/D N/D 434

378.2, 1.46 1.14 +++ N/D N/D 435

378.2, 1.44 1.13 +++ N/D N/D 436

385.1, 2.25 2.58 ++++ N/D +++ 437

374.0, 2.14 2.42 ++++ N/D +++ 438

400.0, 1.90 2.04 ++++ N/D +++ 439

367.1, 2.20 2.47 ++++ N/D +++ 440

367.1, 2.07 2.29 ++++ N/D +++ 441

374.1, 2.07 2.26 ++++ N/D +++ 442

379.1, 1.94 2.19 ++++ N/D +++ 443

364.1, 1.41 1.10 ++++ N/D N/D 444

364.1, 1.33 1.16 ++++ N/D N/D 445

364.1, 1.37 1.10 ++++ N/D N/D 446

475.4, 1.99 2.52 ++++ N/D ++ 447

418.3, 1.54 1.93 ++++ N/D ++ 448

380.1, 1.98 2.06 ++++ N/D +++ 449

375.0, 2.00 2.21 ++++ N/D +++ 450

380.1, 2.01 2.19 ++++ N/D ++ 451

381.0, 1.30 1.48, (7.22) ++++ N/D +++ 452

483.0, 2.83 ++++ N/D +++ 453

467.0, 2.87 ++++ N/D +++ 454

483.0, 2.83 ++++ N/D +++ 455

467.0, 2.87 ++++ N/D +++ 456

525.1, 2.90 ++++ N/D +++ 457

599.2, 3.60 ++++ N/D +++ 458

495.1, 2.77 ++++ N/D +++ 459

425.1, 1.80 ++++ N/D N/D 460

511.1, 3.28 ++++ N/D +++ 461

503.1, 2.66 ++++ N/D +++ 462

275.0, 1.16 1.23, (5.79) ++++ N/D +++ 463

274.0, 1.36 ++++ N/D ++ 464

307.9, 2.09 ++++ N/D ++ 465

352.0, 2.46 357, (17.04) ++++ N/D +++ 466

326.2, 1.66 2.04, (10.20) ++++ N/D ++ 467

360.2, 2.18 2.92, (14.71) ++++ N/D ++ 468

321.2, 1.84 2.35, 11.87 ++++ N/D ++ 469

482.4, 1.70 2.08, (10.76) ++++ N/D N/D 470

417.3, 1.58 1.83, (9.39) ++++ N/D +++ 471

326.3, 1.98 2.53, (13.21) ++++ N/D N/D 472

327.2, 2.21 3.13, (15.01) ++++ N/D +++ 473

326.3, 1.76 2.24, (11.11) +++ N/D N/D 474

327.3, 1.97 269, (12.81) ++++ N/D ++ 475

380.3, 1.49 1.76, (8.70) ++++ N/D +++ 476

395.3, 1.89 ++++ N/D +++ 477

422.3, 2.15 ++++ N/D +++ 478

273.2, 1.55 1.44, (6.80) ++++ N/D ++ 479

307.1, 2.05 2.33, (11.43) ++++ N/D N/D 480

395.3, 1.79 ++++ N/D +++ 481

422.3, 2.10 ++++ N/D ++++ 482

2.73.2, 1.29 1.43, (6.78) ++++ N/D +++ 483

307.2, 1.96 2.58, (12.75) ++++ N/D ++ 484

366.3, 1.39 1.63, (7.75) ++++ N/D +++ 485

505.1, 2.35 14.35 L ++++ N/D N/D 486

487.2, 2.31 13.84 L ++++ N/D N/D 487

427.1, 2.12 11.84 L ++++ N/D N/D 488

544.2, 1.76 1.67 ++++ N/D +++ 489

581.2, 1.82 1.90 ++++ N/D +++ 490

491.1, 1.70 1.59 ++++ N/D N/D 491

547.1, 2.09 11.59 L ++++ N/D ++ 492

492.9, 1.78 2.24 ++++ N/D N/D 493

561.1, 2.20 2.46 ++++ N/D N/D 494

430.2, 1.97 10.65 L ++++ N/D ++ 495

485.0, 2.47 15.16 L ++++ N/D ++ 496

484.1, 2.47 15.14 L ++++ N/D N/D 497

462.4, 1.29 7.09 L ++++ N/D +++ 498

463.2, 1.77 8.92 L +++ N/D N/D 499

463.2, 1.72 8.24 L ++++ N/D +++ 500

428.4, 1.63 10.17 L ++++ N/D +++ 501

462.4, 1.28 1.51 ++++ N/D +++ 502

511.3, 2.08 11.82 L +++ N/D N/D 503

428.2, 1.93 10.1 L ++++ N/D +++ 504

378.2, 1.66 7.72 L +++ N/D N/D 505

378.2, 1.76 8.73 L ++++ N/D +++ 506

447.3, 1.78 8.02 ++++ N/D N/D 507

461.2, 1.8 8.93 ++++ N/D N/D 508

353.1, 2.16 2.34 ++++ N/D +++ 509

476.3, 1.65 7.27 L ++++ N/D +++ 510

351.1, 1.74 1.70 +++ N/D N/D 511

402.2, 1.65 7.51 L ++++ N/D +++ 512

351.2, 1.66 7.85 L ++++ N/D +++ 513

258.2, 1.48 6.33 L ++++ N/D N/D 514

372.2, 1.65 7.49 ++++ N/D +++ 515

359.2, 2.05 11.16 L ++++ N/D ++++ 516

308.2, 1.54 6.71 L ++++ N/D +++ 517

373.2, 0.71 5.93 +++ N/D N/D 518

336.2, 1.61 8.11 L ++++ N/D ++++ 519

383.2, 1.44 2.04 ++++ N/D N/D 520

383.2, 1.53 2.09 ++++ N/D +++ 521

390.1, 1.59 7.15 L ++++ N/D +++ 522

390.1, 1.75 8.62 L ++++ N/D +++ 523

293.1, 1.93 2.20 ++++ N/D +++ 524

489.1, 2.47 ++++ N/D +++ 525

495.2, 2.49 ++++ N/D N/D 526

485.1, 2.90 ++++ N/D +++ 527

459.2, 2.75 ++++ N/D +++ 528

495.2, 2.47 ++++ N/D +++ 529

415.1, 2.06 ++++ N/D +++ 530

413.1, 3.09 ++++ N/D +++ 531

413.1, 3.07 ++++ N/D +++ 532

515.1, 2.74 ++++ N/D +++ 533

481.1, 2.54 +++ N/D N/D 534

497.1, 3.01 ++++ N/D +++ 535

545.1, 3.37 ++++ N/D N/D 536

515.1, 2.79 ++++ N/D N/D 537

481.1, 2.55 ++++ N/D N/D 538

497.1, 2.54 3.00, (15.55) ++++ N/D N/D 539

469.1, 2.56 ++++ N/D +++ 540

489.1, 2.47 ++++ N/D +++ 541

417.0, 1.51 1.84, (8.78) ++++ N/D ++ 542

469.0, 1.76 2.27, (10.99) ++++ N/D N/D 543

481.1, 1.93 2.57, (12.58) ++++ N/D +++ 544

536.9, 2.47 3.38, (17.30) ++++ N/D ++ 545

449.9, 3.44 N/D N/D ++ 546

460.0, 3.00 ++++ N/D +++ 547

484.5, 2.28 3.12, (15.46) +++ N/D N/D 548

427.3, 1.83 2.49, (11.84) ++++ N/D +++ 549

427.3, 1.82 2.51, (11.79) ++++ N/D +++ 550

360.9, 1.56 N/D N/D +++ 551

358.9, 1.63 ++++ N/D +++ 552

558.3, 1.90 ++++ ++++ +++ 553

588.3, 1.92 ++++ ++++ ++++ 554

500.0; 2.46 ++++ N/D +++ 555

514.0; 2.62 +++ N/D N/D

The compounds in Table 1 were synthesized according to Methods 1-30 andExamples 1-35 provided above. PI3K IC₅₀ values and pSer473 Akt EC₅₀values for inhibition of Akt phosphorylation were determined accordingto Biological Methods 1 and 2, respectively. The cellular proliferationEC₅₀ values shown in Table 1 were determined according to BiologicalMethod 3.

Table 1 shows the IC₅₀ and EC₅₀ values of the compounds as determined bythe Biological Methods 1, 2, 3 and 4 as described herein. In Table 1,“+” indicates that the compound had an IC₅₀ or EC₅₀ value of >25 μM;“++” indicates that the compound had an IC₅₀ or EC₅₀ value of <25 μM;“+++” indicates that the compound had an IC₅₀ or EC₅₀ value of >10 μM;and “++++” indicates that the compound had an IC₅₀ or EC₅₀ value of >1μM. An “N/D” in Table 1 indicates that the values were not determined.

Each of the Compounds in Table 1 exhibited IC₅₀ values of less than 10μM with respect to inhibition of PI3K. Many of the Compounds of Table 1exhibited IC₅₀ values of less than 1 μM and even less than 0.1 μM withrespect to inhibition of PI3K. For this reason, each of the compounds isindividually preferred and is preferred as a group. The PI3 kinase alphaIC₅₀ values shown in Table 1 were determined according to the ATPdepletion assay as disclosed herein in Biological Method 1.

Furthermore, many of the compounds of Table 1 exhibited an EC₅₀ valuewith respect inhibition of pSer473 Akt phosphorylation of less than 10μM. Many of those compounds exhibited EC₅₀ values of less than 1 μM andeven less than 0.1 μM with respect to pAkt inhibition. Table 1 shows theEC₅₀ values for inhibition of phosphorylation of pSER473 AKT. The assayswere performed according to the Biological Method 2 described herein.

In addition, many of the compounds of Table 1 were tested to determinetheir inhibitory activity in a cellular proliferation assay according toBiological Method 4. Many of those compounds exhibited EC₅₀ values ofless than 1 μM and even less than 0.1 μM, demonstrating their potentability to inhibit cellular proliferation. Table 1 shows the EC₅₀ valuesfor inhibition of cellular proliferation of an ovarian cancer cell line,A2780/

Biological Method 1 Phosphorylation Assays

Assay 1: Homogenous Solution Phase Assay

Compounds to be tested are dissolved in DMSO and directly distributedinto 384-well flashplates at 1.25 μL per well. To start the reaction, 20μL of 6 nM PI3 kinase are added into each well followed by 20 μL of 400nM ATP containing a trace of radiolabeled ATP and 900 nM1-alpha-phosphatidylinositol (PI). The plates are briefly centrifuged toremove any air gap. The reaction is performed for 15 minutes and thenstopped by the addition of 20 μL of 100 mM EDTA. The stopped reaction isincubated overnight at RT to allow the lipid substrate to bind byhydrophobic interaction to the surface of the flashplate. The liquid inthe wells is then washed away, and the labeled substrate is detectedwith scintillation counting.

Assay 2: One Step Solid Phase Assay

This method is similar to Assay 1 except that the lipid substrate(1-alpha-phosphatidylinositol (PIP)) is first dissolved in a coatingbuffer and incubated on flashplate at room temperature overnight toallow the lipid substrate to bind by hydrophobic interaction to thesurface of the flashplate. Unbound substrate is then washed away. On theday of assay, 20 μL of 6 nM PI3 kinase are added into each well followedby 20 μL of 400 nM ATP containing trace of radiolabeled ATP. Compoundsare added together with enzyme and ATP to the lipid-coated plates. Theplates are briefly centrifuged to remove any air gap. The reaction isperformed for two to three hours. The reaction is stopped by addition of20 μL of 100 mM EDTA or by immediate plate washing. Phosphorylated lipidsubstrate is detected by scintillation counting.

Assay 3: ATP Depletion Assay

Compounds to be tested are dissolved in DMSO and directly distributedinto a black 384-well plate at 1.25 μL per well. To start the reaction,25 μL of 10 nM PI3 kinase and 5 μg/mL 1-alpha-phosphatidylinositol (PI)are added into each well followed by 25 μL of 2 μM ATP. The reaction isperformed until approx 50% of the ATP is depleted, and then stopped bythe addition of 25 μL of KinaseGlo solution purchased from Promega. Thestopped reaction is incubated for 5 minutes and the remaining ATP isthen detected via luminescence.

Biological Method 2 pSer473 Akt Assays to Monitor PI3K Pathway

In this method, an assay for measuring the PI3K-mediated pSer473-Aktstatus after treatment with representative inhibitor compounds of theinvention is described.

A2780 cells were cultured in DMEM supplemented with 10% FBS.L-glutamine, sodium pyruvate, and antibiotics. Cells were plated in thesame medium at a density of 15,000 cells per well into 96 well tissueculture plates, with outside wells vacant, and allowed to adhereovernight.

Test compounds supplied in DMSO were diluted further into DMSO at 500times the desired final concentrations before dilution into culturemedia to 2 times the final concentrations. Equal volumes of 2× compoundswere added to the cells in 96 well plates and incubated at 37° C. forone hour. The media and compounds were then removed, the plates chilledand cells lysed in a lysis buffer (150 mM NaCl, 20 mM Tris pH 7.5, 1 mMEDTA, 1 mM EGTA, 1% Triton X-100) supplemented with phosphatase andprotease inhibitors. After thorough mixing, lysates were transferred toboth pSer473Akt and total Akt assay plates from Meso Scale Discovery(MSD), and incubated overnight with shaking at 4° C. The plates werewashed with 1×MSD wash buffer and the captured analytes detected withsecondary antibodies. After incubation with the secondary antibody atroom temperature for 1-2 hours, the plates were washed again and 1.5×concentration of Read Buffer T (MSD) was added to the wells.

The assays were read on a SECTOR Imager 6000 instrument (Meso ScaleDiscovery). Ratios of the signal from pSer473Akt and total Akt assayswere used to correct for any variability and the percent inhibition ofpSer473Akt from the total signal seen in cells treated with compoundversus DMSO alone was calculated and used to determine EC₅₀ values foreach compound.

Biological Method 3 Pharmacology Target Modulation and Efficacy Study inOvarian Cancer Xenograft Model

A2780 ovarian cancer cells obtained from George Coukos (Fox Chase CancerCenter, University of Pennsylvania, Philadelphia, Pa.) were maintainedin DMEM (Invitrogen, Inc.) supplemented with 10% heat-inactivated fetalbovine serum with 1% glutamine. Cells were propagated as recommended bythe Dr. Coukos and colleagues. Female nu/nu mice (8-12 weeks old, 20-25g, Charles River) were used for all in vivo pharmacology studies. Themice were housed and maintained in accordance with state and federalguidelines for the humane treatment and care of laboratory animals andreceived food and water ad libitum. Cancer cells were harvested frommid-log phase cultures using trypsin-EDTA (Invitrogen, Inc.). Fivemillion cells were subcutaneously injected into the right flank of eachmouse. Compound treatment was initiated when tumor size reached to300-400 mm³ for PK/PD studies and 200-300 mm³ for efficacy studies. Allcompound treatment was administrated orally. Tumor volumes weredetermined by using StudyDirector software.

For in vivo target modulation PK/PD time-course studies, tumor tissueswere resected from individual mice at different time points ranging from30 min to 36 hr after a single dose of compound (60 or 100 mg/kg) orvehicle was administrated orally. For PK/PD dose-dependent studies,tumor-bearing mice were given a single oral dose of compound atdifferent concentrations (10, 30, 60 and 100 mg/kg or vehicle) andtumors were resected at 10 hr or 24 hr after dosing. Blood samples weretaken by cardiac puncture using a syringe primed with heparin sulfate.Resected tumors were snap frozen on dry ice and pulverized using aliquid nitrogen-cooled cryomortar and pestle, and lysed in cold cellextraction buffer (Biosource) containing protease inhibitor tablet(Complete; EDTA-free, Amersham). Supernatants were taken aftercentrifugation of tumor lysates at 300×g for 10 min at 4° C. and theprotein concentration in each supernatant was determined by BCA(BioRad). An equal amount of protein from each tumor lysate was loadedonto 10% Tris-glycine gels (Invitrogen), forsodiumdoceylsulfate-polyacrylamide gel electrophoresis (SDS-PAGE) afterwhich proteins were transferred from the gel onto PVDF membrane.Membranes were probed with antibodies that recognize phosphoAkt^(Ser473)or phosphoAkt^(Thr308) (Cell Signaling) followed by secondary goatanti-rabbit IgG conjugated to HRP(Amersham). Positive bands werevisualized by enhanced chemiluminescence with X-ray film. Similarprocedures were used to determine total AKT in the same tumor lysates toserve as normalization for total protein in each determination. Thedensity of the positive band on the X-ray film was scanned and thetarget modulation for each compound was expressed as percentageinhibition by each compound compared to vehicle treatment. A rank order(<50%, 50-75%, >75%, as compared to vehicle treatment) of targetinhibition is used to present compound target modulation activity.

For efficacy studies, A2780 cancer cells (5×10⁶ in 100 μl of DMEMculture medium) were injected subcutaneously into the right flank ofeach nu/nu mouse. When average tumor sizes reached about 200 mm³, micewere dosed orally daily (q.d.) or twice a day (b.i.d.) at threedifferent compound concentrations (typically at 10, 30 and 100 mg/kg) in100 μl incipient. Tumor growth and animal body weight was measured twiceweekly with daily clinical observation to monitor potential toxicitiesrelated to the treatment. Typically, studies were terminated when tumorsin vehicle-treated group reached 2500 mm³ or adverse clinical effectswere observed. Activation of the PI3K signaling pathway results in thephosphorylation of the downstream signaling molecule Akt at Ser⁴⁷³and/or Thr³⁰⁸. Compound modulation of Akt^(Ser473) phosphorylation wasexamined in A2780 xenograft tumors at time points ranging from 30 min to36 hr after a single compound dose at 60 or 100 mg/kg. Table 2summarizes modulation of AKT^(Ser473) phosphorylation by representativecompounds at 8 hr or 10 hr time points. Percentage inhibition was rankedas <50%, 50-75%, and >75%, as compared to vehicle treatment.

TABLE 2 Modulation of Akt^(Ser473) phosphorylation by representativepyrimidine compounds of the invention. Compound 60 mg/kg 100 mg/kg  91at 8 hr >50% 183 at 8 hr 50-75% 103 at 8 hr <50%  10 at 10 hr >75% >75% 84 at 10 hr 50-75%  76 at 10 hr >75% >75%  66 at 10 hr <50%

Efficacy of Compound 91 was tested in the A2780 tumor xenograft model.Mice bearing A2780 tumors received oral administration of Compound 91twice daily at 10 and 60 mg/kg. Tumor growth inhibition (50%) wasobserved at 60 mg/kg treatment, while at 10 mg/kg no inhibitory activitywas observed (FIG. 1).

The modest tumor growth inhibition by Compound 91 at 60 mg/kg q.d. wasdue to its short-lived target modulation (50% inhibition lasted for 8hr). Therefore, antitumor efficacy of three other compounds (Compound10, Compound 76, and Compound 66), that demonstrated longer inhibitionof Akt^(Ser473) (>50% inhibition>10 hr) in A2780 tumors were evaluatedin A2780 model. Compounds were orally administrated daily when tumorsizes reached to about 200 mm³. Compound 10 demonstrated adose-dependent tumor growth inhibition: 40% at 30 mg/kg, 70% at 60 mg/kgand tumor growth stasis at 100 mg/kg (FIG. 2). A similar dose-dependenttumor growth inhibition was observed with Compound 76 treatment at 30and 60 mg/kg in the A2780 tumor model (FIG. 3) while Compound 84 wasfound to possess weaker antitumor activity (<50% TGI at 60 mg/kg) (FIG.4).

Antitumor activity of Compound 10 was also evaluated at more frequentdosing regimen (b.i.d). As shown in FIG. 5, Compound 10 demonstrated asignificant antitumor activity at 30 mg/kg when orally dosed b.i.d.Notably, tumor growth inhibition at 30 mg/kg b.i.d. was more potent thanwhen dosed on a schedule at an equivalent daily dose (60 mg/kg, FIG. 2).The compounds were well tolerated in this study. This result indicatedthat a sustained but less profound target inhibition (covering the wholedosing period, but with <75% target inhibition) in A2780 tumors byCompound 10 was able to induce significant antitumor efficacy.

While the preferred embodiment of the invention has been illustrated anddescribed, it will be appreciated that various changes can be madetherein without departing from the spirit and scope of the invention.

Biological Method 4 Cellular Proliferation Studies in A2780 Cell

The ability of the compounds of the invention to inhibit cellularproliferation were determined by using Cell Titer Glo, a commerciallyavailable assay from Promega Corporation. A2780 ovarian cancer cellswere seeded in TC treated 96-well plates at a density of 1,000 per wellin DMEM, 10% FBS, 1% Sodium Pyruvate, and 1% Penicillin Streptomycin fora minimum of 2 hrs prior to addition of compound. For each concentrationof test compound, 2 μl (500×) aliquots of compound or 100% DMSO dilutedin 500 μl of culture medium for 2× concentration then diluted 1× on theCells. Cells were incubated for 72 hrs at 37° C., 5% CO₂. After the 72hour incubation, Cell Titer Glo reagent is added to determine number ofviable cells remaining after exposure to the compound, and the EC₅₀value was calculated. The assay was performed according to themanufacturer's instruction (Promega Corporation, Madison, Wis. USA).Each experimental condition was performed in duplicate. The results areprovided in Table 1

1. The compound4-(trifluoromethyl)-5-(2,6-dimorpholinopyrimidin-4-yl)pyridin-2-amine,of structure:

or a pharmaceutically acceptable salt thereof.
 2. A composition,comprising a pharmaceutically acceptable carrier and an amount of acompound of claim 1 effective to inhibit PI3-K activity in a human oranimal subject when administered thereto.
 3. The composition of claim 2further comprising at least one agent for the treatment of cancer. 4.The composition of claim 3, wherein the at least one agent for thetreatment of cancer is vatalanib, imatinib, erlotinib, lapatinib,pertuzumab, trastuzumab, capecitabine, gemcitabine, irinotecan,paclitaxel, cisplatin, carboplatin, fulvestrant, dexamethasone,bevacizumab, docetaxel or gefitinib.
 5. A method for treating ovariancancer in a human or animal subject having ovarian cancer, comprisingadministering to the human or animal subject a composition comprising anamount of a compound of claim 1 effective to inhibit PI3-K activity inthe human or animal subject.
 6. The method of claim 5 further comprisingadministering to the human or animal subject at least one agent for thetreatment of cancer.
 7. The method of claim 6, wherein the at least oneagent for the treatment of cancer is vatalanib, imatinib, erlotinib,lapatinib, pertuzumab, trastuzumab, capecitabine, gemcitabine,irinotecan, paclitaxel, cisplatin, carboplatin, fulvestrant,dexamethasone, bevacizumab, docetaxel or gefitinib.